AmScope 490 Series Operator's Manual

Microscope) 490 Series Compound Microscope

AmScope Proprietary Page 2 of 64 Copyright © 2015 United Scope Inc. DBA AmScope. All rights reserved. Copyright Notice This document, which is in draft form and subject to change without notice, is an unpublished work containing confidential, trade secret information that is proprietary to AmScope, and is protected by laws pertaining to such materials. This document, the information in this document and all rights thereto are the sole and exclusive property of AmScope and are not to be copied, used or disclosed to anyone, in whole or in part, without the express written permission of AmScope. AmScope assumes no liability for any damages incurred, directly or indirectly, from any errors, omissions or discrepancies between the use of this microscope and the information contained in this document. AmScope Purchasing: 949-333-0001 or 1-888-950-2888 FREE (toll free) Customer Service: 949-333-0002 Technical Support: 949-333-0004 Shipping: 949-333-0005 Accounting: 949-333-0008 Fax: 949-333-0007

AmScope Proprietary Page 3 of 64 Contents BEFORE USE ..................................................................................................................................................................... 4 Introduction ................................................................................................................................................................. 4 Precautions ................................................................................................................................................................. 4 1. MICROSCOPE PARTS ..................................................................................................................................................... 5 2. OPERATION ................................................................................................................................................................... 6 2.1 Unpacking ............................................................................................................................................................. 6 2.2 Assembly ............................................................................................................................................................... 7 2.3 Adjusting the View .............................................................................................................................................. 10 2.4 Specimen Set Up .................................................................................................................................................. 11 2.5 Focusing .............................................................................................................................................................. 12 2.6 Using the Trinocular Port ................................................................................................................................... 13 2.7 Attaching a Camera ............................................................................................................................................ 13 2.8 Setting the Stage’s Stop-Limit ............................................................................................................................. 14 2.9 Adjusting Focus Tension ..................................................................................................................................... 14 2.10 Setting the Condenser Lens Adjustment Knob................................................................................................... 15 2.11 Adjusting the Iris ............................................................................................................................................... 15 2.12 Changing a Filter .............................................................................................................................................. 15 2.13 Changing the Halogen Light Bulbs ................................................................................................................... 16 2.14 Changing the Fuse ............................................................................................................................................ 16 2.15 Using Oil Immersion ......................................................................................................................................... 17 2.16 Using Darkfield Condenser Lenses (Optional Accessories) ............................................................................. 19 2.17 Using a Dry Darkfield Condenser Lens (Optional Accessory) ......................................................................... 20 2.18 Using Oil Darkfield Condenser Lens (Optional Accessory) ............................................................................. 21 2.19 Using a Phase Contrast Kit (Optional Accessory) ............................................................................................ 24 2.20 Using a Simple Phase Contrast Kit (Optional Accessory) ................................................................................ 28 2.21 Using a Turret Phase Contrast Kit (Optional Accessory) ................................................................................. 33 2.22 Microscope Maintenance .................................................................................................................................. 38 2.23 Darkfield Condenser Kit & Phase Contrast Kit Maintenance .......................................................................... 39 3. GENERAL SPECIFICATIONS .......................................................................................................................................... 40 3.1 General Model Comparison ................................................................................................................................ 40 3.2 Objectives ............................................................................................................................................................ 41 3.3 Eyepieces ............................................................................................................................................................. 41 3.4 Darkfield Condensers.......................................................................................................................................... 41 3.5 Phase Contrast Kits............................................................................................................................................. 42 4. PARAMETERS .............................................................................................................................................................. 43 4.1 Electrical System ................................................................................................................................................. 43 4.2 Microscope Parameters ...................................................................................................................................... 43 5. RECOMMENDED ACCESSORIES ................................................................................................................................... 44 5.1 Eyepieces ............................................................................................................................................................. 44 5.2 Darkfield Condensers.......................................................................................................................................... 44 5.3 Objective Lenses ................................................................................................................................................. 45 5.4 Phase Contrast Kits............................................................................................................................................. 45 5.5 Cameras and Accessories ................................................................................................................................... 46 5.6 Halogen Light Bulbs and Fuses .......................................................................................................................... 47 5.7 Cases and Bags ................................................................................................................................................... 47 5.8 Stage Warmer ...................................................................................................................................................... 47 5.9 Slides ................................................................................................................................................................... 48 5.10 Cleaners ............................................................................................................................................................ 48 5.11 Books ................................................................................................................................................................. 48 6. TROUBLESHOOTING .................................................................................................................................................... 49 6.1 Optical Issues ...................................................................................................................................................... 49 6.2 Mechanical Issues ............................................................................................................................................... 50 6.3 Electrical Issues .................................................................................................................................................. 51 7. GENERAL MICROSCOPY GUIDE ................................................................................................................................... 52

AmScope Proprietary Page 4 of 64 Before Use Introduction Congratulations on the purchase of your new AmScope Microscope! This manual is designed for 490 series microscopes. Though the microscope’s body and magnification levels may vary depending on your series model number, the 490 series functions remain the same. Please take a few minutes to familiarize yourself with the features and functions of your new microscope. If you want more information on microscope, parts, and accessories, please visit our website at: www.AmScope.com We recommend that you study this manual thoroughly before operating the microscope and that you keep it on hand for future reference. If you have additional questions or need assistance, please send us an email at: [email protected] Please include the microscope model number in your email so that we can identify your model and provide immediate help. Precautions 1. As the microscope is a precision instrument, always handle it with care, avoiding impact or abrupt movement during transportation. Do not shake the package. 2. Do not place the microscope in direct sunlight or in high heat. Keep it indoors in a dry and clean place with temperatures between 32°-100° F (0°-40°C), maximum relative humidity: 85% 3. Avoid touching the lenses on the objective and the eyepiece so that oil and dirt from your fingerprints do not obstruct your view. 4. Before turning the power on, make sure that the power supply voltage is consistent with the voltage of your microscope.

AmScope Proprietary Page 6 of 64 2. Operation 2.1 Unpacking 2.1.1 Unpack the Box 1. Very carefully slide the Styrofoam container out of the cardboard carton. 2. Lay the Styrofoam container on its side. Make sure the side labeled up is up. 3. Remove the tape. 4. Carefully open the Styrofoam container, avoid dropping and damaging the optical items. 2.1.2 Verify Packing List Check the packing list to ensure that you’re received all items: � One Microscope Body � One Binocular (B490) or One Trinocular (T490) Rotating Gemel Head � One Adjustable Trinocular Photo Port (for T490 only) � Four Objectives (4x, 10x, 40x, 100x) � Pair of 10X Widefield Eyepieces � Pair of 16X Widefield Eyepieces (for -A models only) � Pair of 20X Widefield Eyepieces (for -B models only) � Pair of 25X Widefield Eyepieces (for -C models only) � Three Color Filters (Blue, Green, & Yellow), (for non-LED models only) � One Bottle of Immersion Oil � One 6V/20W Spare Halogen Bulb (for 20W models only) � One 6V/30W Spare Halogen Bulb (for 30W models only) � One Spare Fuse � One Dust Cover Note: LED models do not have a spare bulb, as the LED bulbs do not need replacing.

AmScope Proprietary Page 7 of 64 2.2 Assembly 2.2.1 Carefully Remove Microscope from Box Remove the microscope body from the box and remove the plastic protective covering. The body of the microscope is composed of the base, the stage, the arm, and the nosepiece. 2.2.2 Loosen the Head Lock Screw Loosen the metal knob located directly above the nosepiece (the head lock screw) completely. 2.2.3 Install the Microscope Head Carefully take the microscope head out of the plastic bag, making sure not to touch glass. Place the bottom side of the microscope head (flat, circular side down) into the circular opening on top of the arm (just above the nosepiece and lock screw). Tighten the head lock screw to secure the head in place.

AmScope Proprietary Page 8 of 64 2.2.4 Install the Trinocular Port and C-Mount (For Trinocular Microscopes Only) Unscrew and remove the photo port cap from the top side of the microscope head. Screw in the Trinocular port If your camera has a c-mount, insert the c-mount adapter onto the Trinocular port. If your camera does not have a c-mount, you can skip this step. 2.2.5 Insert Eyepieces Remove the eyetube caps. Place the desired eyepieces into the empty eye/ocular tubes. Be sure to avoid touching the lenses.

AmScope Proprietary Page 9 of 64 2.2.6 Insert Objective Lenses Unpack the objective lenses. Screw the objectives into the microscope nosepiece. Start with the lowest magnification, and in order of magnification, insert each lens, without touching the glass. 2.2.7 Plug it in, turn it on, and adjust the dimmer if more light is needed. Plug in the microscope and turn it on. If no light emerges from the light source, adjust the dimmer knob on the side of the base.

AmScope Proprietary Page 10 of 64 2.3 Adjusting the View 2.3.1 Choose forward or reverse viewing The microscope head rotates to allow for forward or reverse viewing. There is no difference as far as the optics; it depends on what’s comfortable. In forward position you are looking over the stage, in reverse position you are looking at the stage. Once you’ve chosen your preferred microscope head position, lock the head-lock screw. 2.3.2 Focus the distance between your eyes. (The interpupillary distance, the distance between your eye pupils.) With both eyes open, look into the eyepieces. Adjust the interpupillary distance by holding the eyetubes and rotating the tubes either towards or away from each other until only one circle of light is seen by both eyes.

AmScope Proprietary Page 11 of 64 2.4 Specimen Set Up 2.4.1 Prepare the Specimen Place the specimen to be studied on a glass slide (or use a prepared slide, sold separately). 2.4.2 Secure the Slide Place the slide on the stage, holding it snugly in place with the metal slide holders (clips) of the mechanical stage. 2.4.3 Center the Specimen Using the mechanical stage controls, center the slide over the stage opening, lining it up with the light and the objective lens. The top ring moves the stage forward and backward. The bottom ring moves the stage from left to right 2.4.4 Adjust the dimmer if more light is needed. To adjust the illumination, slowly turn the dimmer on the right side of the base until the desired intensity of light is achieved.

AmScope Proprietary Page 12 of 64 2.5 Focusing 2.5.1 Choose an Objective Lens Turn the nosepiece to choose an objective. It is easiest to use the lowest magnification first (4X objective) to locate and focus on the specimen. As you move up in magnification you will need to refocus the image a little each time. 2.5.2 Parfocal Under the diopters on the eyepieces, there are numbered rings. Set these rings to 0 and the image should remain in focus for the entire magnification range. This will enable you to stay more in focus when you add a camera or switch objective lenses. 2.5.3 Focus one eye at time 1. Look with one eye through the right eyetube. Or, if you have two diopters, select either eyepiece. 2. Close your other eye. 3. Focus the image using the coarse focusing knob. Adjust the height of the stage until the sample comes into clear focus. Note: You can loosen the limit-stop knob (located on the inside of the coarse focusing knob on the left side of the microscope) in order to give yourself the full range of motion. 2.5.4 Focus your other eye 1. Close the first eye. Open the other one. 2. Look with the second eye through the eyepiece. 3. Turn the diopter until the image is clear through the side as well. Do not turn the focusing knob when you are adjusting the diopter or you will have difficulty focusing properly. 2.5.5 Re-center Specimen and Re-focus When you change magnification levels, you may need to re-center the specimen. When you change magnification, you will need to re-focus.

AmScope Proprietary Page 13 of 64 2.6 Using the Trinocular Port The AmScope T490 models are uniquely designed so that you can view the image through the eyepieces and the Trinocular port simultaneously, as well as fine tune the focus of the camera with a C-mount focus adjustment. You do not need an adapter to attach your AmScope camera to the Trinocular port; however you may need one if you have a non-AmScope camera. Our photo port is a 23mm size. 2.7 Attaching a Camera Most AmScope cameras come with a reduction lens. Reduction lenses align the magnification of the microscope’s image with the capacity of the digital camera’s sensor. The specimen’s image seen through the magnified objective lens is very large. The reduction lens renders the image in a size compatible with the smaller size of the digital camera’s sensor. 1. If your camera has a 23mm mounting size, you do not need the C-mount adapter. If installed, remove the C- mount from the top of the Trinocular port by loosening the screw. 2. Place the camera (mounted on a reduction lens) directly into the Trinocular port. It should slide in without issue. 3. Focus through the Trinocular port, by turning the middle portion of the tube. 4. Open up your AmScope Digital Camera Solution software on your computer and let it connect to your microscope. If your camera needs a C-mount adapter, attach the adapter to your camera and screw the adapter into the Trinocular port. Plug one end of the cord directly into the camera and the USB end into your computer.

AmScope Proprietary Page 14 of 64 2.8 Setting the Stage’s Stop-Limit 1. Unlock the stop-limit on the stage. 2. Adjust the stage to the desired maximum height. 3. Lock the stop-limit. This allows you to limit the movement of the stage from the bottom of the range up to the point it is set at. 4. To reset it, unlock the stage and reset the stage to the new height. If no limit is desired, simply unlock the stop-limit. 2.9 Adjusting Focus Tension 1. To adjust the tension of the focusing knobs, first locate the black ridged tension knob on the inside of the coarse focusing knob. 2. To decrease tension, rotate the adjustment forward, towards the stage (counterclockwise). To increase, rotate away from the stage (clockwise). Note: If your stage is slipping down after setting the focus, you need to increase the tension.

AmScope Proprietary Page 15 of 64 2.10 Setting the Condenser Lens Adjustment Knob The condenser adjustment knob raises and lowers the condenser’s distance to the base lens, varying light delivery. 2.11 Adjusting the Iris By changing the aperture (hole size) of the iris/diaphragm of the condenser, you can adjust the background brightness. Adjust the aperture of the iris diaphragm using the iris adjustment slider located directly under the stage. 2.12 Changing a Filter 1. At the bottom of the condenser lens, there is a small ring that slides out. Swivel the ring as far out as possible. 2. Insert the preferred colored glass filter. 3. Slide the ring back into place. Note: The filter holder is placed in from the factory in a manner in which it swings out and hits the arm of the microscope (backwards). If this happens, simply grip the condenser assembly and rotate it. It may take a small amount of force to rotate it, but after doing so, you will be able to swing the filter holder out towards the front of the unit for easier operation.

AmScope Proprietary Page 16 of 64 2.13 Changing the Halogen Light Bulbs 1. Turn off and unplug the microscope. 2. Wait until the bulb cools down. 3. Do not touch the bulb directly, the oil from your skin will damage the bulb. Use a cloth or paper to handle. (Microscope Base) 1. Be sure to remove the eyepieces from the unit before turning the microscope over to prevent them from falling and breaking. 2. Carefully turn the microscope on its side, exposing the base. 3. Loosen the door screw on the bottom of the microscope and pull the door open. 4. Remove the old bulb and replace with a new one. 5. Close door and tighten the screws of the door. 2.14 Changing the Fuse (Microscope Base) 1. Turn off and unplug the microscope. 2. Be sure to remove the eyepieces from the unit before turning the microscope over to prevent them from falling and breaking. 3. Carefully turn the microscope on its side, exposing the base. 4. Using a small screwdriver, unscrew the cover and replace the fuse. 5. Screw the fuse cover back in.

AmScope Proprietary Page 17 of 64 2.15 Using Oil Immersion When using the 100X objective, a drop of immersion oil should be placed between the cover slip and the objective to minimize distortion caused by air. 1. Place the slide on the stage and center the specimen. 2. Close the condenser’s aperture iris and ensure that the specimen focused and centered as possible. Increase light intensity once the specimen is clearly illuminated. 3. Move the objectives out of the way to enable access to the slide. Place a drop of immersion oil directly onto the slide’s cover slip, not directly on the specimen; a cover slip MUST be used. 4. Release the stage stop limit. 5. Lower your oil immersion lens (100X objective) into position so that the front lens is immersed in the oil. If your lens is not immersed, use the coarse focusing knobs to adjust the height of the stage to the proper levels. The oil lens should be immersed in oil, but not touching the slide.

AmScope Proprietary Page 18 of 64 6. Adjust the focusing knob, watch for the field of view to come into focus. This is best seen when the iris of the condenser is smaller than the field of the objective. Use the fine focusing knob to make adjustments. 7. Re-engage the stage stop limit. 8. When the specimen is in focus, adjust the iris to open just larger than the condenser’s diameter. 9. After using the oil immersion lens, you will need to clean the lens and specimen before the oil dries. Note: Please be careful when using the stage stop limit. The stage limit stop is designed to prevent impact between objective and slide, so when it is off you can damage the microscope or the slide.

AmScope Proprietary Page 19 of 64 2.16 Using Darkfield Condenser Lenses (Optional Accessories) Darkfield Illumination is a lighting method for increasing contrast. A darkfield condenser lens produces bright images against a dark background for low contrast specimens. The dry darkfield condenser is for low power magnifications, such as with the 4X, 10X or 40X objective lenses, with numeric apertures of .65 or less. The oil darkfield condenser is for high power magnifications, the 40X and 100X objective lenses, with numeric apertures up to 1.25 or less. 2.16.1 Installing a Darkfield Condenser Lens 1. Use the condenser lens adjustment knob to lower the condenser so you have easy access to the condenser knob. 2. Loosen the condenser lock screw and replace the condenser lens. 3. Install the darkfield condenser lens and tighten the condenser lock thumb screw on the condenser holder. When choosing a darkfield condenser, be sure the condenser’s numerical aperture is larger than the numerical aperture of the objective lens. If the objective lens aperture is larger than the aperture of the darkfield condenser, direct light will be transmitted and a darkfield will not be achieved.

AmScope Proprietary Page 20 of 64 2.17 Using a Dry Darkfield Condenser Lens (Optional Accessory) 2.17.1 Using the Dry Darkfield Condenser 1. The first step is centering the condenser. Move the 10X objective lens into the light path. 2. Remove an eyepiece and look through the empty eye tube, to see the condenser’s image directly. 3. Use the condenser lens adjustment knob to adjust the height of the condenser until the opaque disc is slightly smaller than the entire image circle, leaving a ring of light. 4. Turn the condenser’s centering screws to move the dark spot to the center. 5. Replace the eyepieces. 6. Raise the condenser to the top of its range, just short of the slide on the stage. The is the optimal position for darkfield viewing, but can be adjusted later, if necessary. 7. Place a slide on the stage and focus. Adjust the height of the stage up or down slightly for optimal viewing, but the condenser lens should be relatively close to the slide. 8. If you are using a variable numerical aperture objective lens, adjust the ring at the bottom to fine tune the darkfield.

AmScope Proprietary Page 21 of 64 2.18 Using Oil Darkfield Condenser Lens (Optional Accessory) The condenser works with 100X oil objective and low power objectives (4X, 10X, etc) as well. Immersion oil is needed on top of the condenser lens for darkfield observation. Immersion oil is needed between the 100X oil objective and slide. 2.18.1 Using the Oil Darkfield Condenser 1. The first step is centering the condenser. Move the 40X objective lens into the light path. 2. Remove an eyepiece and look through the empty eye tube, to see the condenser’s image directly. 3. Use the condenser lens adjustment knob to adjust the height of the condenser until the opaque disc is slightly smaller than the entire image circle, leaving a ring of light. Note: The dark spot has a fuzzy edge and will become larger when the condenser gets closers to the objective. The distance between the top of the condenser and the objective is about 8mm, when the dark spot shows in the field of view. 4. Turn the condenser’s centering screws to move the dark spot to the center. 5. Replace the eyepieces. 6. Raise the condenser to the top of its range, just short of the slide on the stage. The is the optimal position for darkfield viewing, but can be adjusted later, if necessary. 7. Adjust the dimmer so the lamp is at maximum brightness. 8. Place a slide on the stage and focus. Adjust the height of the stage up or down slightly for optimal viewing, but the condenser lens should be relatively close to the slide.

AmScope Proprietary Page 22 of 64 2.18.2 Using Oil Immersion 1. Before you install the oil darkfield condenser lens, apply a drop of immersion oil on the top lens of condenser. If you’ve already installed it, remove the condenser to apply the drop of oil to the lens, and re-install it. You might need to re-center the condenser. 2. Place the slide on the stage. 3. Raise the condenser and let the oil drop contact the underside of the slide. If air bubbles exist in the oil, clean the oil from the condenser lens and bottom of slide and repeat the procedures. 4. Apply a drop of immersion oil to the slide’s coverslip. 5. Carefully raise the stage until the oil makes contact with the objective lens. 6. Adjust the iris ring to get proper brightness and contrast of view field. 7. If you are using a variable numerical aperture objective lens, adjust the ring at the bottom to fine tune the darkfield.

AmScope Proprietary Page 23 of 64 2.18.3 Cleaning the Oil Darkfield Condenser 1. When using oil immersion, once you have finished using the oil darkfield lens, you will need to clean the lens and specimen of the oil before it dries. 2. You can wipe the lens with a soft nonabrasive cloth to remove the oil. If you want to use a cleaner, you can use cigarette lighter fluid to safely remove the oil, as it will break it down. Do not use alcohol based solvents, as they dissolve the cement used to in assembly of the lens (thus breaking your objective). 3. The specimen itself can be discarded, or if desired, you could clean it up and store it for later use. The specimen may also be left with the oil on it, as you will still need to apply more for use later, however it may dry up and cloud your specimen. Cleaning is recommended.

AmScope Proprietary Page 24 of 64 2.19 Using a Phase Contrast Kit (Optional Accessory) Brightfield Phase Contrast Some specimens may offer little in the way of color-variance or opacity and can appear essentially invisible when using basic, transmitted light. Phase-contrast is a technique used in microscopy to enhance the contrast of an image, when observing a specimen which offers little contrast in normal bright-field conditions. Light waves have several measurable components, the most-common of which are length, frequency and amplitude. While length and frequency translate into the color of light we see, amplitude translates into brightness. One component that we do not perceive is phase, which can be described as “how much of the wave’s cycle has elapsed at a given time.” If you imagine light as a small dot (a photon), moving along the cyclical arcs of a light wave, the phase is current position of the photon, relative to the wave’s entire cycle.

AmScope Proprietary Page 25 of 64 As waves of light move through space, if their cycles move in a perfect parallel and are at the same point in their cycles, they are in- phase. If some of the light waves hit an obstacle and the phases of the waves are altered, so they are no longer in parallel and at the same point in their cycles, they are out-of- phase. When a light wave passes through any medium, the medium can affect the light wave’s length, amplitude, frequency and phase. There are shifts in phase, due to changes in light’s speed. As the speed of light changes when hitting or missing obstacles, like a specimen, the rays are bent, or refracted. The amount of refraction when a light wave hits an obstacle is measured as its refractive index. It’s this refractive index which creates the variations in phase as light passes through a specimen. Even though a specimen may appear transparent, as long as its refractive index is different from the baseline index of air, and the specimen has thickness, then there will be changes in phase to any light which passes through the specimen.

AmScope Proprietary Page 26 of 64 A phase contrast system incorporates two components to segregate light, so out-of- phase waves can be realigned. The two components are the annulus in the condenser and the phase ring in the objective lens. The condenser’s annulus separates light into surround light and diffracted light. Light passing directly through the annulus creates a hollow cone called surround light. This light passes directly through the optical system and is intercepted by the phase ring in the objective lens. Light diffracted by the edges of the annulus passes more centrally through the specimen in the objective lens. As the light collectively passes through the objective lens, it is further segregated by the phase ring. The phase ring shifts the phase of the surround light either positively or negatively so the waves are in-phase. Since human sight is incapable of perceiving light phase, the aim of phase contrast is to translate the disparities in phase into perceivable variations in brightness. There are several techniques available for increasing the contrast between the specimen and the background, each tailored for a particular purpose. Phase-contrast does so by manipulating the phase of light, and is useful for observing the inner mechanics of cellular structures, especially live cells, without the need for staining.

AmScope Proprietary Page 27 of 64 AmScope’s PCS Simple Phase Contrast Kit includes three phase-contrast objective lenses, three phase contrast condenser lenses, and a centering telescope. AmScope’s PCT Turret Phase Contrast Kit includes four phase-contrast objective lenses, a condenser turret, and a centering telescope. The turret has built-in annuli, designed to work with each of the lenses, as well as a brightfield iris.

AmScope Proprietary Page 28 of 64 2.20 Using a Simple Phase Contrast Kit (Optional Accessory) 2.20.1 Unpacking the Simple Phase Contrast Kit 1. Take the aluminum case containing your kit out of the cardboard carton. Remove the tape and open the container carefully so as to avoid dropping and damaging the optical items. You may need to use the key taped onto the box. Check carefully to ensure that all parts and accessories are intact. Note: You must match finite tube-length kits to finite tube-length microscopes and infinity-corrected units to infinity-corrected microscopes for optimal results. 2. Check the packing list to ensure that you’re received all the items. All units should come with: � 10X, 40X, & 100X Phase Contrast Condenser Lenses � 10X, 40X, & 100X Phase Contrast Objective Lenses � Condenser Assembly � Centering Telescope Lens � Green filter

AmScope Proprietary Page 29 of 64 2.20.2 Focus the Specimen 1. Once you have installed the phase contrast kit, you will need to calibrate and center the light perfectly for optimal results. You will initially calibrate with the Abbe brightfield condenser. 2. Place a slide on the microscope stage. Make sure the specimen is covered with a cover slip (a good example for a phase contrast specimen is human saliva). Clean any excess material from the slide. 3. Make sure your Abbe brightfield condenser is installed. 4. Focus the specimen using the 10X objective. Adjust the iris ring on the condenser so the light field is just larger than the field of view in the microscope. The condenser itself should be moved to the normal height, which is close to the bottom of the stage of the microscope (the top of the range of the condenser). 2.20.3 Installing a Simple Phase Contrast Condenser 1. Loosen the thumbscrew and remove the existing Abbe brightfield condenser from your microscope. 2. Replace the brightfield objective on nosepiece with the 10X phase contrast objective. 3. Screw the 10X condenser ring plate onto the condenser assembly. Install the ring plate and assembly on the microscope’s condenser ring.

AmScope Proprietary Page 30 of 64 4. Change the objective lens to the 10X phase contrast lens. Objective lenses designed for phase contrast will be marked “Ph”, this denotes the objective lens has a built-in phase ring. Note: there are 3 phase contrast objectives: 10X, 40X and 100X, and there are 3 condenser ring plates: 10X, 40X and 100X. The corresponding objective and ring plate must work together. For example, the 10X phase contrast objective must work with the 10X condenser ring plate. 2.20.4 Calibrating the Phase Contrast Objective Lens and Condenser Lens 1. Remove one eyepiece from the microscope eyetube and insert the centering telescope. 2. Turn the microscope’s light on and look through the telescope lens. 3. While looking through the eyepiece, extend the eyepiece by rotating the top counter clockwise until an image of the objective’s phase ring and the condenser’s annulus are clearly in focus. Since they are both part of the same conjugate image plane, they can be simultaneously in focus.

AmScope Proprietary Page 31 of 64 4. If the bright ring is still obscure, raise or lower the condenser by adjusting the condenser focusing knob (or the microscope focusing knob if necessary) until the bright ring is in focus and the dark ring is visible. 5. If the two ring images are not positioned so one is inside the other, adjust the centering screws on the condenser ring plate until the two rings center in parallel. The annulus’ bright ring of light should overlap the dark grey ring, which is the objective’s phase ring. 6. Place the green filter in the filter holder or directly onto the center of the base lens of your microscope (where the light emerges from). Remove the green filter if doing darkfield microscopy. 7. Remove the centering telescope and replace the eyepiece. Note: Once aligned, repeat the process for each magnification.

AmScope Proprietary Page 32 of 64 2.20.5 Perform the Phase Contrast Observation After you center the condenser, you can perform the phase contrast observation the same way as a normal brightfield microscope. Note: When changing to another phase contrast objective and corresponding condenser ring plate, the focusing and centering of bright ring and dark ring should be repeated following the procedures above. Tips: 1. Make the illumination as bright as possible. 2. The thinner the specimen, the better the image.

AmScope Proprietary Page 33 of 64 2.21 Using a Turret Phase Contrast Kit (Optional Accessory) 2.19.1 Unpacking the Turret Phase Contrast Kit 1. Take the aluminum case containing your kit out of the cardboard carton. Remove the tape and open the container carefully so as to avoid dropping and damaging the optical items. You may need to use the key taped onto the box. Check carefully to ensure that all parts and accessories are intact. Note: You must match finite tube-length kits to finite tube-length microscopes and infinity-corrected units to infinity-corrected microscopes for optimal results. 2. Check the packing list to ensure that you’re received all the items. All units should come with: � Turret with 10X, 20X, 40X &100X Phase Contrast Condensers � 10X, 20X, 40X & 100X Phase Contrast Objectives � Centering Telescope Lens � Green filter � Small flathead screwdriver

AmScope Proprietary Page 34 of 64 2.21.2 Installing the Turret Phase Contrast Kit 1. Loosen the lock screw and remove the Abbe brightfield condenser from the holder. 2. Insert the phase contrast condenser turret into the condenser holder and tighten the lock screw with the screw driver. Due to the weight of the turret, it’s best to use the screw driver to securely fasten the turret to the condenser holder. 2.21.3 Focusing the Turret Phase Contrast Kit 1. Rotate the microscope’s nosepiece to the 10X phase contrast objective lens. Objective lenses designed for phase contrast will be marked “Ph”, this denotes the objective lens has a built-in phase ring. Note: There are 4 phase contrast objectives: 10X, 20X, 40X and 100X, and there are 5 settings on the condenser’s turret ring: 10, 20, 40, 100 and B. The numbers on the turret ring must always correspond with objective’s numbers. For example, when using a 10X phase contrast objective the condenser’s turret ring setting must be 10. The B setting will make the phase contrast condenser work as a conventional brightfield condenser.

AmScope Proprietary Page 35 of 64 2. Place a slide on the microscope stage. Make sure the specimen is covered with a cover slip (a good example for a phase contrast specimen is human saliva). Clean any excess material from the slide. 3. Turn the phase contrast condenser’s turret to the B setting. Focus the specimen. Before aligning the phase contrast lens and condenser, you will need to bring the specimen into focus with the brightfield setting. 4. Once the specimen is in focus with the brightfield setting, rotate the condenser’s turret ring so it is set to 10. 2.21.4 Calibrating the Turret Phase Contrast Objectives and Condenser Lenses The turret has a built-in annulus for each objective magnification. Once the turret and objective lens are set with matching numbers, they will need to be aligned. You will need to do this again each time you change the objective lens. Each annulus in the turret is easy to reposition by using a finger. 1. Remove one eyepiece from the microscope eyetube and insert the centering telescope. 2. Turn the microscope’s light on and look through the telescope lens.

AmScope Proprietary Page 36 of 64 3. While looking through the telescope lens, extend the eyepiece by rotating the top counter clockwise. 4. Keep rotating the telescope lens until an image of the objective’s phase ring and the condenser’s annulus are clearly in focus. 5. By slowly moving the annulus on the underside of the turret, now in-line with the condenser’s lens, the annulus’ bright ring of light should overlap the dark grey ring, which is the objective’s phase ring 6. If the bright ring is still obscure, raise or lower the condenser by adjusting the condenser focusing knob (or the microscope focusing knob if necessary) until the bright ring is in focus and the dark ring is visible. 7. Place the green filter in the filter holder or directly onto the center of the base lens of your microscope (where the light emerges from). Remove the green filter if doing darkfield microscopy. 8. Remove the centering telescope and replace the eyepiece. Note: Once aligned, repeat the process for each magnification.

AmScope Proprietary Page 37 of 64 2.21.5 Perform the Phase Contrast Observation After you center the ring plate, you can perform the phase contrast observation the same way as a normal brightfield microscope. Note: When changing to another phase contrast objective and corresponding condenser ring plate, the focusing and centering of bright ring and dark ring should be repeated following the procedures above. Tips: 1. Make the illumination as bright as possible. 2. The thinner the specimen, the better the image.

AmScope Proprietary Page 38 of 64 2.22 Microscope Maintenance 2.22.1 Handle with Care As the microscope and all accessories are precision instruments, always handle them with care, avoiding impact or abrupt movement during transportation. Do not shake the package. 2.22.2 No Sun/Heat Do not place the microscope and accessories in direct sunlight or in high heat. Keep the microscope and all accessories indoors in a dry and clean place with temperatures between 32°-100° F (0°-40°C), maximum relative humidity: 85%. 2.22.3 No Dust Always keep the microscope covered by the dust cover when not in use. The glass will attract dust, obscuring your view. Make sure to keep it in a dry and clean place in order to prevent rust. 2.22.4 Keep Glass Clean All glass surfaces must always be kept clean. Oil and dirt from your fingerprints will obstruct your view. Fine dust on the optical surface should be blown off using a hand blower or gently wiped off with a soft lens tissue. 2.22.5 Use an optical lens cleaner. Carefully wipe off oil or fingerprints on lens surfaces using a tissue moistened with a small amount of 3:7 alcohol to ether mixture. 2.22.6 Do not use lens cleaner on the non-glass parts of the microscope. Do not use the optical lens cleaning solution to wipe the surfaces of the other components of the microscope. All other parts, especially those made of plastic, should be cleaned with a mild detergent. 2.22.7 Do not touch the glass. Avoid touching the lenses on the objective, the eyepiece and all the accessories with glass lenses.

AmScope Proprietary Page 39 of 64 2.22.8 Do not take apart the microscope. Do not assemble or disassemble the microscope’s electrical components yourself. . 2.23 Darkfield Condenser Kit & Phase Contrast Kit Maintenance 2.23.1 Secure Setting Ensure that the microscope is located on a smooth, level and firm surface. 2.23.2 Do Not Take Apart Do not attempt to disassemble any components. 2.23.3 Keep Kits Clean Keep the kits clean; remove dirt and debris regularly. Accumulated dirt on outer surfaces should be cleaned with a damp cloth. More persistent dirt should be removed using a mild soap solution. Do not use organic solvents for cleansing. 2.23.4 Keep Lens Clean The outer surface of the optics should be inspected and cleaned periodically using an air stream from an air bulb. If dirt remains on the optical surface, use a soft cloth or cotton swab dampened with a lens cleaning solution (available at the camera stores). All optical lenses should be swabbed using a circular motion. A small amount of absorbent cotton wound on the end of a tapered stick makes a useful tool for cleaning recessed optical surfaces. Avoid using an excessive amount of solvents as this may cause problems with optical coatings or cemented optics or the flowing solvent may pick up grease making cleaning more difficult. 2.23.5 Cool and Dry Storage Store the instrument in a cool, dry environment. Put the condenser back in the storage box when not in use.

AmScope Proprietary Page 40 of 64 3. General Specifications 3.1 General Model Comparison PARTS SPECIFICATIONS B490 T490 B490- 30W B490- LED T490- 30W T490- LED WF Eyepiece WF10X/18mm ● ● ● ● ● ● WF10X/18mm w/ Pointer WF10X/18mm w/ Reticle WF16X/13mm ● (if -A) ● (if -A) ● (if -A) ● (if -A) ● (if -A) ● (if -A) WF20X/10mm ● (if -B) ● (if -B) ● (if -B) ● (if -B) ● (if -B) ● (if -B) WF25X/9mm ● (if -C) ● (if -C) ● (if -C) ● (if -C) ● (if -C) ● (if -C) DIN Achromatic Objectives 4X/0.10 ● ● ● ● ● ● 10X/0.25 ● ● ● ● ● ● 40X (spring)/0.65 ● ● ● ● ● ● 100X (spring, oil)/1.25 ● ● ● ● ● ● 30 Degree Viewing Head Binocular Sliding 360 Degree Swiveling ● ● ● Trinocular Sliding 360 Degree Swiveling ● ● ● Trinocular Port C-Mount Adjustable Photo port ● ● ● 23mm Adjustable Photo Port ● ● ● Simultaneous Viewing ● ● ● Focusing Nosepiece Coaxial Focusing System ● ● ● ● ● ● Quadruple Nosepiece ● ● ● ● ● ● Stage 130mm x 140mm Mechanical Stage ● ● ● ● ● ● Movement Range 50mm x 76mm ● ● ● ● ● ● Condenser Abbe, NA= 1.25 ● ● ● ● ● ● Diaphragm Iris Diaphragm ● ● ● ● ● ● Illumination Halogen Light w/ Dimmer ● ● ● ● LED Light w/ Dimmer ● ● Lamp 6V/20W ● ● 6V/30W ● ● LED ● ● Filter Blue/Yellow/Green ● ● ● ●

AmScope Proprietary Page 41 of 64 3.2 Objectives Type Model Number Magnification Numerical Aperture (N.A.) Rating Medium Magnification Marks (Color Ring) DIN Achromatic Objective with Associated N.A. Ratings (195mm) A4X 4X 0.10 Air Red A10X 10X 0.25 Air Yellow A40X 40X 0.65 Air Light Blue A60X 60X 0.85 Air Dark Blue A100X 100X 1.25 Cedar Oil White Plan Objective (195mm) PA4X Plan 4X 0.10 Air Red PA10X Plan 10X 0.25 Air Yellow PA40X Plan 40X 0.65 Air Light Blue PA100X Plan 100X 1.25 Cedar Oil White 3.3 Eyepieces Eyepieces Magnification 5X 10X 15X 16X 20X 25X Field of View Φ18 Φ18 Φ13 Φ15 Φ11 Φ9 3.4 Darkfield Condensers Specifications Dry Darkfield Condenser Oil Darkfield Condenser Model DK-DRY100 DK-OIL100 Numerical Aperture (N.A.) Rating 0.7 – 0.9 1.36 – 1.25 Mounting Size (Diameter) 37mm 37mm

AmScope Proprietary Page 42 of 64 3.5 Phase Contrast Kits Specifications Simple Phase Contrast Kit Turret Phase Contrast Kit Model PCS PCT Phase Contrast Objective Achromatic 10X/0.25, 160/0.17 with built-in phase plate Plan phase contrast 10X/0.25, 160/0.17 Plan phase contrast 20X/0.40, 160/0.17 Achromatic 40X/0.65, 160/0.17 with built-in phase plate, spring Plan phase contrast 40X/0.65, 160/0.17 Achromatic 100X/1.25, 160/0.17 with built-in phase plate, spring, oil Plan phase contrast 100X/1.25, 160/0.17, Oil Numerical Aperture (N.A.) Rating 1.25 1.25 Turret Ring Plates 1 For 10X phase contrast objective 10 for 10X phase contrast objective 20 for 20X phase contrast objective 1 For 40X phase contrast objective 40 for 40X phase contrast objective 1 For 100X phase contrast objective 100 for 100X phase contrast objective Centering Telescope Focusing adjustable Focusing adjustable

AmScope Proprietary Page 43 of 64 4. Parameters 4.1 Electrical System There are two options for electrical systems for this series of microscope. The light source is dependent on which model you have, but can either be a 6V/20W halogen, a 6V/30W halogen, or an LED system. 1. 100V-120V power supply: 100V-120V ±10%, 50/60Hz. 2. 220V-240V power supply: 220V-240V ±10%, 50/60Hz. All units come standard as 110V units unless an upgrade to a 220V system is requested. Upgrade fee is dependent on which unit is purchased. 4.2 Microscope Parameters Magnification -A Model: 40x-1600x -B Model: 40x-2000x -C Model: 40x-2500x Field of View Φ0.8mm~Φ4.5mm Mechanical Tube Length 160mm Object to Primary Image Distance 195mm Fine Focusing Sensitivity 0.002mm

AmScope Proprietary Page 44 of 64 5. Recommended Accessories (Purchase separately. Please visit www.amscope.com and search with SKU #, for more information.) 5.1 Eyepieces 5X Eyepieces 20X Eyepieces 25X Eyepieces SKU: EP5X23 SKU: EP20X23 SKU: EP25X23 10X w/ Pointer 10X w/ Reticle SKU: EP10X23P SKU: EP10X23R 5.2 Darkfield Condensers Dry Darkfield Condenser Oil Darkfield Condenser SKU: DK-DRY100 SKU: DK-OIL100

AmScope Proprietary Page 45 of 64 5.3 Objective Lenses Achromatic 2X 5X 20X 60X SKU: A2X SKU: A5X SKU: A20X SKU: A60X Plan Achromatic Plan 4X Plan 10X Plan 40X Plan 100X SKU: PA4X SKU: PA10X SKU: PA40X SKU: PA100X 5.4 Phase Contrast Kits Simple Turret SKU: PCS SKU: PCT

AmScope Proprietary Page 46 of 64 5.5 Cameras and Accessories Cameras To capture images, video, or view live display on a computer (PC/Mac OS X). Real- Time Live Video Microscope Digital Camera, Reduction Lens, High Sensitivity Sensor and Deluxe Calibration Kit. 350K Pixel 1.3 Mega Pixel 3 Mega Pixel 5 Mega Pixel 8 Mega Pixel 9 Mega Pixel 10 Mega Pixel SKU: SKU: SKU: SKU: SKU: SKU: SKU: MU035 MU130 MU300 MU500 MU800 MU900 MU1000 Calibration Micrometer Calibrate the camera software for on-screen measurements. SKU: MR400 Charge-Coupled Device Television CCD Digital (VGA, Trinocular Only) CCD TV/Video (Trinocular Only) 1920x1080 Full HD HDMI (Trinocular Only) SKU: CCD-MT SKU: CCD-NP SKU: HD1080 To view live display on a computer monitor (VGA). To view live display on a television (RCA). To view live display on a computer monitor (VGA).

AmScope Proprietary Page 47 of 64 5.6 Halogen Light Bulbs and Fuses 6V 20W Halogen Bulb 6V 30W Halogen Bulb Fuses For Microscopes SKU: BH- 6V20W SKU: BH- 6V30W SKU: FS-M T1AL 220V 5.7 Cases and Bags Aluminum Case Hard Wood Case Vinyl Bag SKU: AC-B400 SKU: WC-B400 SKU: CB-B490 5.8 Stage Warmer Stage Warmer TCS-100 Microscope Temperature Control Stage Slide Warmer

AmScope Proprietary Page 48 of 64 5.9 Slides Prepared 25 Slides 50 Slides 100 Slides 200 Slides SKU: PS25W SKU: PS50 SKU: PS100A SKU: PS200 Blank 50 Blank Slides 100 Blank Slides 144 Blank Slides 300 Blank Slides SKU: BS-50P-C SKU: BS-100P- 100S-22 SKU: BS-144P- 200S-22 SKU: BS-300P- 300S 5.10 Cleaners Sparkle Microscope 3 in 1 Professional 3 in 1 Cleaning Kit Optical Lens Cleaner Cleaning Kit for for Microscopes, SKU: CLS Microscopes, Cameras Cameras and and Laptops LCD Screens SKU: CK-I SKU: CK-II 5.11 Books Complete Book of the Microscope The Ultimate Guide to Your Microscope The World of the Microscope (What to look for) Ages 8-Adult (Science and Experiments) Ages 8-Adult Ages 8-12 SKU: BK-CM SKU: BK-UG SKU: BK-WM

AmScope Proprietary Page 49 of 64 6. Troubleshooting 6.1 Optical Issues SYMPTOM SYMPTOM CAUSE REMEDY OPTICAL ISSUES One side of the field of view is darker The nosepiece is misaligned. Turn the nosepiece until it clicks into place. Stains, dust, or dirt has accumulated on the objectives or eyepieces. Clean all lenses with lens cleaner or a lint free non-abrasive cloth. Obstructions are observed in the field of view Stains, dust, or dirt has accumulated on the specimen. Clean the slide or use a new specimen if sample is destroyed. Stains, dust, or dirt has accumulated on the objective, eyepieces, or Barlow lens. Clean the lens. Unclear Image There is no cover slip on the slide. Add a cover slip. The objectives are designed for use with a 0.17mm cover slip, so it is a requirement to use one for proper images. The cover slip is not standard sized. Replace the cover slip with the appropriate 0.17mm thickness slip. The immersion oil has accumulated on the dry objective. Thoroughly clean the objective lens with lens cleaner or a lint free non-abrasive cloth No immersion oil is used with the 100X objective. Use immersion oil for better clarity and resolution. Used wrong oil. Use standard cedar wood oil. The aperture is not open to an appropriate diameter. Adjust the aperture to have the light just larger than the size of the condenser. Stains, dust, or dirt has accumulated on the inlet of the head. Clean the lens with lens cleaner or a non- abrasive lint free cloth, as well as spray with compressed air. The condenser is not in the right position. Adjust condenser height to the top of the travel range, and then adjust down to focus image. Initial focus was at too high a magnification. Start focusing with the lowest objective and then switch to the higher ones. The slide is upside down The slide thickness at higher magnifications will impact clarity if the slide is upside down. The color of the image is not accurate The brightness adjustment knob is not in the right position. Adjust the brightness knob to a higher or lower setting for color clarity. No filter is used or filter is in use. Remove color filter if natural light is desired, or insert desired filter.

AmScope Proprietary Page 50 of 64 SYMPTOM CAUSE REMEDY OPTICAL ISSUES One side of the field of view is dark or the image moves while focusing The specimen slide is not fixed. Secure the slide to the stage with clips. The nosepiece is not in the right position. Turn the nosepiece until it clicks into place. The field of view is not bright enough The iris diaphragm is too small. Adjust the iris diaphragm to allow the light to be just larger than the condenser. The condenser is not in the right position. Adjust condenser height to the top of the travel range, and then adjust down to focus image. Stains, dust or dirt has accumulated on the condenser, objective, eyepieces, or base lens. Clean the lens with lens cleaner or a non-abrasive lint free cloth. 6.2 Mechanical Issues SYMPTOM CAUSE REMEDY MECHANICAL ISSUES Unable to move the slide smoothly. The slide is not secured correctly. Adjust the slide to use the stage clips and secure the sample. The mechanical stage is not properly secured. Tighten the mechanical stage screws to better secure the stage. The objective touches the cover slip. The cover slip is not standard sized. Replace the cover slip with the appropriate 0.17mm thickness slip. The limit-stop is set too high or not engaged. Be careful to avoid contact between objective and the slide when the limit stop is not engaged (unless using the 100X objective with oil). To re-engage, focus the sample, then lock the limit stop into place to set maximum height at a safe but usable distance. Focus-knob does not turn. The tension knob is too tight. Loosen it by adjusting the tension ring inside the coarse focus knob counterclockwise (close to the arm of the microscope on the left of the microscope). Stage declines by itself. The tension knob is too loose. Tighten it by adjusting the tension ring inside the coarse focus knob clockwise (close to the arm of the microscope on the left of the microscope). The coarse focusing knob won’t raise the stage. Limit-stop is engaged. Disengage the limit stop on the left side of the microscope inside the coarse focusing knob. The fine focusing knob won’t raise the stage. Limit-stop is engaged. Disengage the limit stop on the left side of the microscope inside the coarse focusing knob.

AmScope Proprietary Page 51 of 64 6.3 Electrical Issues SYMPTOM CAUSE REMEDY ELECTRICAL ISSUES The bulb/light source flickers. The bulb is close to burning out. Replace the bulb. This unit uses our BH- 6V20W with our 20w unit or our BH- 6V30W for the 30W unit. The microscope does not light up. The microscope is unplugged. Insert the plug into the wall socket to achieve electrical illumination. The bulb is not inserted correctly. Check the bulb by unscrewing the base (remove eyepieces first to prevent falling out) door and ensuring that the bulb is inserted. The bulb burned out. Replace the bulb. This unit uses our BH- 6V20W with our 20w unit or our BH- 6V30W for the 30W unit. The fuse burned out. Replace with fuse on the bottom of the microscope. The fuse burns out frequently. The voltage is too high. Use the correct power supply (110v if 110v unit, 220v if 220v unit), or get a voltage adapter to convert to the proper electrical system. The bulb burns out frequently. The voltage is too high. Use the correct power supply (110v if 110v unit, 220v if 220v unit), or get a voltage adapter to convert to the proper electrical system. Used wrong bulb. Use the correct wattage bulb for the unit. Using a higher wattage than it is rated for can damage your unit (melt components with additional heat), so please be sure to use the correct one. Damage from incorrect usage is not covered under warranty.

AmScope Proprietary Page 52 of 64 7. General Microscopy Guide Microscopes come in a wide variety of types with many different features. Each AmScope model is designed for specific uses and specific users. This glossary will help illustrate what the variations mean and why they are useful. Compound and Stereo Microscopes Using a compound or a stereo microscope depends on the specimen being studied. Compound microscopes are best for smaller transparent specimens, like slides and biological subjects. Compound microscopes show a two-dimensional image of the specimen (usually reversed and upside-down) The common magnification range of our compound microscopes is between 40X-1000X, could be up to 2500X. You need a minimum of 400X to study cell structure. Stereo microscopes are best for larger specimens you cannot see through. Stereo microscopes show a 3D image. Three dimensional imaging is perfect for performing dissections, repairing circuit boards, studying fossils and gems or examining any specimen where you want to use your hands. The magnification range of our stereo microscopes is between 2X to 225X.

AmScope Proprietary Page 53 of 64 Base Lens Base lenses direct the light source towards the specimen. Binocular Head-Rotating A rotating binocular head is designed for maximum flexibility in viewing options. Binocular microscope heads allow for the easy adjustment of the interpupillary distance (the distance between the eyes), by moving the eyepieces toward or away from each other... The rotating feature allows for either forward or reverse view, depending on whether you’d rather look at the specimen or over it. C-Mount C-Mounts are adapters with a standard thread for mounting a lens to a camera. C-Mounts fit onto a Trinocular port. Condenser Lens A condenser lens concentrates light on a specimen and increases the resolution. An adjustable iris controls the diameter of the beam of light entering the lens system. Abbe Condenser lenses are specially designed to mount under the stage.

AmScope Proprietary Page 54 of 64 Condenser Lens Adjustment Knob The Condenser Lens Adjustment Knob changes the distance between the light condenser and the base lens. This allows you to control the concentration of the light hitting your slide. Darkfield Condenser Lens A darkfield condenser lens produces bright images against a dark background for low contrast biological specimens. With brightfield lenses, a solid beam of light illuminates the objective lens. Darkfield condenser lenses block the center of the beam of light to produce a hollow cone of light. Light does not enter the objective lens directly. The specimen scatters light, which then enters the objective lens. Brightfield Darkfield Dimmer Dimmers control the amount of light escaping from the base lens. DIN/JIS DIN is the Deutsches Institut für Normung, the German Institute for Standardization, an international standards organization determining the "standard" for many types of technology. JIS is the Japanese Industrial Standard, specifying the standards used for industrial activities in Japan.

AmScope Proprietary Page 55 of 64 Diopters Small rings on the eyetubes, used to focus ocular lenses (eyepieces). Eye Guards Eye Guards fit over the eyepieces, they are for comfort and to protect the glass in the eyepiece. Eyepieces Eyepieces are also called ocular lenses. Eyepieces come in many magnifications and you replace them by swapping them out of the eyetubes. The eyepieces magnify the specimen further from the first magnification through the objective lens. Each eyepiece is marked with 2 numbers. The first number is the magnification and the second is the field of view. For example, an eyepiece marked WF10/20 has a magnification of 10X and a field of view of 20mm. Eyetubes Eyetubes are also called ocular tubes. Eyetubes house the eyepieces and camera adapters. You change eyepeices by removing them from the eyetubes and replacing them with the new ones.

AmScope Proprietary Page 56 of 64 Field of View and Working Distance Field of view is how much of the specimen you can see through the eyepiece. The linear field of view of the eyepiece is divided by the magnification of the objective. The higher the magnification, the smaller the field of view. Working distance is the distance between the bottom of objective lens and the stage. You change the working distance when you use the coarse focusing knob. Together, working distance and field of view determine how much of the specimen you see and how closely. Filters Optical filters selectively transmit light of different wavelengths. Blue filters compensate for low light levels in a brightfield application. (Like dry layer blood analysis.) With low light, you see a yellowing effect. The blue filter brings the light back to a more natural white light. Green filters optimize the performance of achromatic and plan achromatic objectives. When using phase contrast the best images are often produced in green light. Yellow filters compensate for blue images. The yellow filter brings the light back to a more natural white light.

AmScope Proprietary Page 57 of 64 Focusing Knobs Coarse Focus: This is the large knob on the side of the microscope that moves the objective lens up and down. It is used in conjunction with the fine focus. Do not use this knob with the 100X objective lens. Fine Focus: The outer knob, used to fine-tune the focus of a specimen in conjunction with the coarse focus. Head Lock Screw The head lock screw secures the microscope head to the microscope. Loosen the head to rotate between forward and reverse mounting. Tighten the screw to lock the microscope head position into place. Head Orientation Microscope head orientation is for user preference only; it makes no difference in imaging. Forward Reverse

AmScope Proprietary Page 58 of 64 Head Types Monocular Monocular microscope heads have one eyepiece. Binocular Binocular microscope heads have two eyepieces. Trinocular Trinocular microscope heads have two eyepieces and a photo port for a camera. This allows you to look at a specimen with a camera without removing an eyepiece. Multi-head Multi head microscopes are specially designed for three-person observation. Ideal for teaching and/or training purposes. It has a binocular head and two monocular heads. Simul-Focal Simul-Focal heads mitigate the amount of re-focusing needed when you change your view from the eyepieces to the monitor. Perfect for lectures, teaching demonstrations, clinical examinations and laboratory applications.

AmScope Proprietary Page 59 of 64 Immersion Oil A special oil used with the 100X objective to concentrate the light and increase the resolution of the image. At high magnifications the field of view is so small that very little light passes. The light refracts, scatters and produces a dark image. A drop of oil is placed on the slide’s cover slip and the objective is lowered until it touches the oil. The oil will match the specimen’s light refracting index to the lens, adding more light to the specimen, which improves resolution and clarity. Interpupillary Distance Interpupillary Distance is the distance between your eyes. AmScope uses 2 types of systems to change interpupillary distance. Siedentopf heads adjust the interpupillary distance by rotating the eyetubes around a central axis. A Siedentopf head changes the interpupillary distance without changing the tube distance. Jentsch heads adjust the interpupillary distance by moving the eyetubes closer together or farther apart in a linear fashion. The tube length changes when changing interpupillary distance, so the user compensates by adjusting the focus of the eyepieces to correspond to the new interpupillary distance. Iris or Diaphragm The diaphragm or iris is located under the stage and is an apparatus that can be adjusted to vary the intensity, and size, of the cone of light that is projected through the slide. The iris adjustment slider allows you to adjust the opening of the iris. Lenses Microscopes have a two lens system, objective and ocular. The Objective Lens is the lens closest to the specimen or object. The Ocular Lens (Eyepiece) is the lens closest to the eye.

AmScope Proprietary Page 60 of 64 Light Bulbs-Caution 1. Turn off the microscope when not in use to preserve the life of the bulb. 2. Never touch a light bulb directly, especially when it’s turned on. a. The light bulbs can get hot enough to burn skin. Wait until the bulb has cooled completely before handling it. b. The oil from your skin will create a hot spot on the bulb which will damage the glass, dramatically shortening the life of the bulb. c. When changing light bulbs cover skin with cloth or paper to handle the bulb. This will protect both you and the bulb. 3. Hot bulbs will not damage the microscope. AmScope’s microscopes are designed specifically to handle the heat output of the model’s bulb. Lighting-Types Tungsten Tungsten light bulbs are the most economical, providing a reliable source of light. They burn hotter than the other bulbs, but are the best option for certain microscopes. Fluorescent Fluorescent bulbs burn cooler and brighter than tungsten bulbs. The bulbs are more expensive than tungsten, but last longer. LED LED light bulbs burn the coolest of all the bulbs and last the longest. The bulbs are also the most expensive. Depending on use, LED lights should last about 40,000 hours. Halogen Halogen light bulbs produce the most amount of light. The light is very white and concentrated; the bulbs can get very hot and must be cooled regularly. Depending on use, halogen light bulbs can last anywhere from 1 week to 6 months. Fiber Optic Fiber optical lighting uses a system of flexible transparent fibers made of plastic or glass. The light is transmitted between the two ends of the fiber, allowing for greater illumination in confined spaces.

AmScope Proprietary Page 61 of 64 Limit Stop Knob The limit stop is designed to prevent impact between objective and slide. You may loosen the limit-stop knob in order to give yourself the full range of motion for fine tuning the focus when using the 40X and 100X objective lenses. When the limit stop is off you will be able to damage the microscope or the glass slide. For safety, engage the limit stop once you have it close or in contact with the objective (if using oil contact is required). Magnification Microscopes have a two lens system. Total Magnification is the power of two lenses multiplied together. E.g.: (10X Eyepieces) x (4X Objective) = 40X Total Magnification Mechanical Stage A Mechanical Stage is a flat mechanism that sits on top of the stage and allows the viewer to move a specimen small distances - a task that is otherwise difficult at higher magnifications. Most mechanical stages are equipped with an X and Y axis so the viewer can see how far the slide has moved. Mechanical Stage Control The mechanical stage control moves the stage, mechanically moving the slide along an X and Y axis for optimal positioning. The top ring moves the stage forward and backward. The bottom ring moves the stage from left to right. Nosepiece This circular structure is where the different objective lenses are screwed in. To change the magnification power, rotate the turret.

AmScope Proprietary Page 62 of 64 Numerical Aperture (N.A.) N.A. is a rating for the resolution of the objective lens. The N.A. rating ranks the objective lens’ ability to capture light and show fine detail. Lens with larger numerical apertures capture finer details than lenses with smaller numerical apertures. Generally, lenses with larger numerical apertures create a brighter image, but the depth of field will be shallower. N.A. Rating Objective Lens Average Numerical Aperture 4X 0.1 10X 0.25 40X 0.65 100X 1.25 Parcentered When changing objectives, the image of the specimen stays centered. Most compound microscopes are parcentered. Parfocal When changing objectives, the image of the specimen stays in focus without needing to adjust the coarse focusing knob. Not all compound microscopes are parfocal. Phase Contrast Many biological specimens are virtually transparent when viewed with a typical compound microscope. Phase contrast microscopy is a method that collects light from specific incidents (phases) of light only to allow better contrast in viewing transparent specimens. Phase Contrast improves visibility by manipulating direct and diffracted light to produce greater contrast without losing resolution. The major benefit is the ability to study living cells in their natural state instead of killing them with stain. Brightfield Phase Contrast

AmScope Proprietary Page 63 of 64 Phase Contrast Condenser Lenses A specific condenser with a ring printed on it, used for phase contrast. The magnification of the objective must match the magnification of the condenser. Condensers are used to concentrate the light from a transmitted light on a sample for illumination and viewing. Phase Contrast Condenser Assembly For phase contrast kits without a turret, the condenser assembly is the mount for the condenser lenses. Phase Contrast Objective A specific objective with a ring printed on it that is used for phase contrast. The objective is the bottom lens that determines the resolution of the image in the microscope, and is one of the two lenses that compound to make total magnification. Phase Contrast Turret The phase contrast condensers are mounted on a rotating turret much like the objectives on a microscope are. The turret also has brightfield and darkfield condensers.

AmScope Proprietary Page 64 of 64 Telescope Lens-Centering A lens that telescopes in one way or another, used in an eyepiece slot to assist in centering the light in phase contrast microscopy. Tension Knob The large, innermost ring on the focusing knob. Adjusts the tension of the focusing knobs. Trinocular Port A trinocular microscope has three viewing ports. There are two eyepieces and a third port for photography and video. The trinocular port is a 23mm adjustable photo port and uses a C-Mount adapter.