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How to use the Microtome Instructuions - Alisha Quinn

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How to use the Microtome Instructuions - Alisha Quinn

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1. Remove any excess wax around the paraffin block to ensure the block will sit

securely in the microtome chuck for even sections of wax to be cut.

2. Pull the leaver of the microtome forward and place the paraffin wax block into

the chuck. Ensure the hand wheel is locked when placing the block into the

chuck. Check if the paraffin block is held securely in the chuck before cutting.

3. Pull the blade guard down and place a disposable blade into the blade holder by

sliding it in carefully to prevent getting cut. When the microtome is not being

used ensure the blade guard is placed over the blade and the hand wheel is

locked.

4. Put the microtome in trim mode, unlock the hand wheel and move the blade

towards the paraffin block until they engage. Rotate the hand wheel

approximately 20 times to trim the excess wax at the front of the block until the

tissue surface is exposed. The paraffin wax should now have a smooth surface

ready for thin sectioning.

5. Remove any wax debris from the stage with a brush to avoid crosscontamination. Place the block on ice for a few moments to avoid crumbling of

the wax when sectioning.

6. Change the setting of the microtome to feed mode, approximately 3mm thick.

7. Rotate the hand wheel slow and steady to create a thin, continuous ribbon

section of tissue. When the tissue begins to hang over the stage use a forceps to

grab the end.

8. Once the tissue section is long enough, take the ribbon off the stage carefully

with a forceps at both ends to avoid breaking the ribbon. Place the ribbon section

on a warm flotation bath. The ribbon section will float on the surface of the water

bath.

9. Label a glass slide with the tissue type, date and patient details.

10.Get the glass slide and place it into the water bath at 90 degrees. Touch the slide

off the tissue section and pull the slide up and out of the water. Place the slide in

an oven to dry.

11.Clean any excess wax off the stage of the microtome with a brush. Place the

blade guard up and ensure the hand wheel is locked. Clear any wax debris from

the flotation bath to avoid cross-contamination between tissue samples.

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1. clean the excess wax off the block by using a blunt scraper such as a spatula. This is done to prevent the block from wobbling as this leads to uneven sections.

2. mount the block to the microtome by pulling the lever forward and ensure it does not move.

3. insert a new disposable bade after the block has been mounted and clamp it in place.

4. put the blade guard up to prevent cutting when not in use and lock the hand wheel to stop the chuck from moving up and down.

5. use the left hand wheel to position the block until the face of the block is just touching the blade

6. use the trimming key in order to face the block by trimming away some of the excess wax that’s on the surface.

7.inspect the surface of the block for excess wax by engaging the lock on the hand wheel and putting on the blade guard before removing the block from the chuck

8. cool the block in order to ensure that flat sections are obtained

9. brush away excess wax on the blade of the microtome to prevent cross contamination of samples

10. re-mount the cooled block and check its position

11.cut the block using the right hand wheel. Gently encourage the sections being produced using a forceps

12. assess the quality of the section being produced. If clumping occurs it means the block is still too warm and will need the be cooled down for an additional 5 minutes

13. once 3-4 good sections have been produced, transfer the ribbon to a water bath using the forceps and a brush

14. mount the sections to a glass slide avoiding a scooping motion. The slide should be placed straight down into the water and brought to the edge of the section and then brought up.

15. remove unwanted sections from the water bath using tissue to prevent cross contamination

16. the slides should be labelled immediately after mounting with your name, the date and the name of the tissue. Place the slides in the oven to dry.

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Microtome Instructions for use - Connor Hogan

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Microtome Instructions for use - Connor Hogan

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1.    Firstly, clean the excess wax off the outside of the cassette with a scraper as it can cause the block to wobble during.

2.    Pull the leaver forward and insert the block into the chuck, make sure the block is not moving, make sure the blade has not been inserted yet ideally when performing this step.

3.    Insert a new disposable blade and place the blade guard over it to avoid cutting yourself when the microtome is not in use and lock the hand wheel when not in use.

4.    Using the left-hand wheel move the entire chuck forward until the face of the block is just touching the blade.

5.    Use the trimming key to face the block which trims away some of the wax on the surface of the block, rotating the wheel 20 times.

6.    Check the block to see if any excess wax remains on it, ensure the wheel has been locked and the blade guard is up before removing.

7.    Cool the block to ensure flat sections are obtained as at room temperature the sections will crumple easily.

8.    While the block is cooling, clean the microtome with a brush to avoid cross-contamination.

9.    Re-mount the cool block and check the position of block again.

10.  Assess the section quality by checking for crumpling and knife marks.

11.  Using the right-hand wheel the sections can now be cut and a forceps can be used as a handle to encourage the sections away from the blade.

12. Further cool the block if necessary if clumping still occurs.

13.  Cut until 3 or 4 good sections have been cut and cradle the sections to the water bath using a forceps and a scalpel/brush and lay it down gently

14.  Take a glass slide, place it straight down into the water bath, bring it the edge and pull straight up.

15. Remove any unwanted sections from the bath using tissue. This will stop cross contamination from occurring.

16. Label the slide using your name, the date and the name of the tissue.

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How to use a Microtome- Greta Grimailaite

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How to use a Microtome- Greta Grimailaite

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1. Clean any wax that is left on the sides using a blunt object, off the cassette to prevent getting uneven sections from the block wobbling on the microtome

2. To mount the block into place, pull the lever that’s in the middle forward and insert the block, ensuring that it is secure and that it won’t fall out.

3. Insert a blade carefully if there is not one there by sliding it into place, using a pencil to fully get the blade into its spot. Clamp the blade in place and immediately put the blade guard up when not in use.

4. Ensure that hand wheel is locked,  and the blade guard is up when not in use

5. Use the left-hand wheel to bring the block forward and to position it for cutting.

6. When the block is positioned in front of the wheel, use the right-hand wheel to move the block up and down while hold the trimming key to trim the wax.

7. Put the blade guard back up and check the block to see if enough wax has been removed. Keep trimming until the entire tissue section is showing through.

8. Cool down the block to prevent the thin sections from crumbling when cutting them.

9. While the block is cooling down, clean the microtome to prevent contamination using a brush in an upward motion to prevent the blade from cutting the brush hairs

10. Remount the cooled block and check the position, adjusting if needed to.

11. When the block is in place, use the right-hand wheel to cut thin sections of the sample. If the sections are clumping, cool down the block and try cutting again. Ensure the blade guard is back up and that the wheel is locked.

12. Once 3-4 good sections have been cut, gently move the sections to a water bath

13. To mount the sections, use a glass slide and dip it into the water bath holding it vertically. Bring the side to the edge of the section and pull the glass slide straight up. Do not scoop the sections.

14. Remove unwanted sections from the water bath using tissue.

15. Label the slides immediately with a name, date and the tissue type.

16. Brush off excess wax off the stage and bring the blade guard back up and ensure the hand wheel is locked.

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Instructions on how to use the Microtome-Lucy Dervan

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Instructions on how to use the Microtome-Lucy Dervan

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1.      Turn on the safety lock on the right hand side of the microtome.

2.      Wipe off any excess wax that is on the microtome.

3.      Place the paraffin wax embedded tissue into the tissue in the centre of the microtome.

4.      Ensure that the right side of the tissue section is exposed to the blade. (You will need to open the tab at the top of the tissue holder before the tissue section can be placed inside).

5.      Once the tissue section is placed inside the tissue holder, ensure the microtome is set to trim.

6.      Remove the blade shield.

7.      Use the up and down arrows on the left corner of the microtome to adjust the tissue section to the blade.

8.      The tissue section should be close enough to the blade to ensure a fine section of tissue will be cut. (If the tissue section is too thick, this can result in the tissue embedded wax falling off of the cassette).

9.        Once the blade is adjusted appropriately, move either handles on the left or right-hand side of the microtome 360°C until the tissue is at the edge of the wax.

10.   Press the button on the left hand side handle of the microtome to switch the microtome from trim to cut.

11.   Using the cut setting, start moving either handles on the left or right-hand side of the microtome 360°C until thin tissue sections are cut.

12.   Use a forceps to remove the tissue section from the blade.

13.   Placed the tissue section in the water bath by first moving the tissue section away from you, then bring the tissues section back to you before placing the tissue section in the water bath.

14.   Stabilise the tissue section in the water bath by dragging the edge of the tissue section to the side of the water bath.

15.   Place a glass slide vertically in the water bath and bring it up to the tissue section in the water bath.

16.   Touch the slide against the tissue section and lift the glass slide up quickly out of the water bath.

17.   Label the glass slide.

18.   Place the slide in the dryer and leave to dry until staining is required.

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1.      Clean off excess wax from the edges of the parafilm block using a blunt scraper. This step ensures the block will not wobble.

2.      Pull the lever and mount the cassette onto the chuck, ensuring the lock is in place so the block will not move.

3.      Insert a disposable blade on the blade plate, ensure the blade guard is up when the blade is not in use to reduce the risk of injury. Additionally, ensure the handwheel lock is on to prevent further risks when the microtome is not in use.

4.      Select the trim function on the desired setting. Unlock the left handwheel and move the wheel so that the blade encounters the block. Trim off excess wax at the front of the block until the tissue is exposed.

5.      Check progress by removing the block from the chuck safely by ensuring the handwheel is locked and the blade guard is up.

6.      Cool the block in ice to ensure high quality cutting such as flat sections with no crumbling.

7.      Brush upwards and clean away any excess wax around the blade plate to prevent any cross contamination between samples.

8.      Remount the block back onto the chuck and check the position to ensure the block is stable.

9.      Change the cutting function to feed mode and select the desired thickness.

10.  Begin to cut thin sections by using the right-hand wheel. Remove the ribbon using a forceps to a water bath when 3-4 sections have been cut. Gently place the ribbon on top of the water.

11.  Mount the section onto a glass slide by placing the slide into the water bath vertically next to the ribbon and pull the slide upwards avoiding a scoping motion.

12.  Use a tissue to remove unwanted tissue samples from the water bath to prevent cross contamination.

13.  Label the glass slide with your initials, date, and sample type.

14.  Place the glass slide into a drying oven

15.  Ensure all equipment is clean, handwheels are locked, and blade guard is up before leaving the microtome station

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1. Clean the block and remove any excess wax from the edges to ensure the block will not wobble and avoids uneven cut sections. A spatula can be used as a blunt edge to remove the excess wax.

2. Before insertion of the blade, pull the lever forwards and insert the block. It is important to make sure that the block is steady and in a fixed position.

3. Insert the blade and avoid contact with the sharp edges. As soon as the blade has been inserted, put up the blade guard.

4. Check safety measures: Ensure the blade guard is up and the handwheel lock should be locked when not cutting. Otherwise, the chuck would move and if the blade guard is not up, it is possible to accidentally cut your hand on the blade.

5. Position the block so that the face of the block is just touching the blade.

6. Use trimming setting on microtome to face the block and trim away the wax on the surface. Remove the block and examine to check that the wax over the face of the specimen has been removed. This can be completed by holding the block in the light and ensuring that the tissue is completely shining through.

7. Cool down the block in order to avoid crumpling of sections. In the meantime, clean down the microtome. Re-mount the cooled block and check the position of the chuck.

8. Take a section and assess the quality of the section for crumpling, knife marks, thickness. Collect sections with a forceps at the end of the section and gently hold the sections upwards, without pulling the sections. If there is any clumping occurring, cool down the block further. Remount, reposition and repeat the process to check for the quality of the section.

9. Transfer sections to a water bath by cradling the section using the forceps and a brush. Gently lower it onto the water.

10. Mount sections by taking a glass slide, putting the glass slide straight down, moving it to the edge and pull straight up. Remove any excess sections in the water bath using a piece of tissue. Lay the tissue across the water and drag it. This reduces the risk of cross-contamination.

11. Label the mounted slides immediately. Must be labelled with your name, date and name of the tissue. Always label after mounting to ensure the identity of the section on the slide is known. While waiting for blocks to cool between mounting, change the blade. Blades go into the sharps bin.

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1. Remove any excess wax from the block to prevent it from wobbling that would cause uneven sections

2. Pull the lever forward and insert the paraffin block containing the tissue and ensure it is secure

3. If necessary, pull the blade guard down to insert a new blade and clamp it in place. Put the blade guard back up and lock the handwheel until you are ready to use the microtome.

4. Use the left handwheel to move the block towards the blade until the face of the block is just touching it

5. Set the right handwheel to the correct cutting width and trim the block by turning the handwheel until the tissue is fully exposed

6. Cool the block and clean excess wax from the microtome to ensure clean sections

7. Re-mount the block and use the right handwheel to get a few sections of the tissue

8. Use forceps to lift the sections and inspect their quality

9. Transfer sections to a water bath and hold a glass slide upright and pull up to gently mount the sections onto it

10. Remove any unwanted sections from the water bath with a clean tissue

11. Label slides with your name, the date and type of tissue

12. If the blade appears dull remove it and dispose of it in a sharps bin

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1. Clean excess wax from around the outside using a blunt scraper, as the excess wax can cause the cassette to wobble, giving uneven sections.

2. Pull the lever forward and insert the block.

3. Insert the blade, clamp it in place and then insert the blade guard. The guard should always be in place while not cutting. The hand wheel should also be locked while not cutting, this is done to prevent the chuck from moving and pushing your hand towards the blade if inserting a cassette.

4. Use the left-hand wheel to move the chuck forward or backward so that the face of the block is just touching the blade.

5. When in position, remove the blade guard and use the trimming key to face block by rotating the right-hand wheel to move the block up and down to remove excess wax to expose the sample.

6. Replace the blade guard and lock the hand wheel and remove cassette to check if all excess wax has been removed. If not, repeat step 5 until the excess wax is trimmed off and the tissue is completely showing.

7. Must cool down the block as at room temperatures the sections that come off are prone to crumpling. This can be done by placing it on ice or a cooling block.

8. Clean down the area to prevent contamination.

9. Once cooled, reinsert the cassette into the chuck and reposition so that the front of the block is at the edge of the blade, Remove the blade guard and lock.

10. Remove a few of the front sections.

11. Use sections ans a handle, grab them, and gently pull them away without tearing them. If the sections are clumping, re-cool the block again on the cooling plate for approximately 5 minutes.

12. Repeat step 11. Try to get around 3 sections that look usable and gently pull the sections.

13. Move the second to a warm water bath and lay them out flat.

14. To mount on slides, insert the slide vertically and move towards the slide. Once the section touches the slide and pull upwards.

15. Label slides with your name, the date and the tissue type.

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Instructions on how to cut a good section using a microtome-McGeorge

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Instructions on how to cut a good section using a microtome-McGeorge

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1.      Clean the block to remove excess wax using a blunt scraper, which could cause wobbling during the cutting, leading to uneven sections.

2.      Mount the block into the chuck by pulling the lever forward and ensuring the block is secure.

3.       Insert the blade by sliding it into the holder and clamp it in place.

4.      Put up the blade guard, which should always be in place when the microtome is not in use. The hand wheel should be locked to avoid the chuck moving while manipulating the block.

5.      Use the left-hand wheel to move the chuck forward to get the face of the block touching the blade.

6.      Turn the microtome to the trim function and face the block to remove the wax on the surface of the tissue.

7.      Engage the wheel lock and blade guard before removing the block for observation of the tissue.

8.      Cool the block in ice or on a cold plate. This prevents rippling in the section caused by the wax melting.

9.      Clean the stage to prevent cross-contamination of tissue samples.

10.   Place the block back into the chuck, ensuring that the wheel lock is engaged and the blade guard is up.

11.   Check the position of the block and move it to the appropriate position for cutting.

12.   Collect sections with forceps, gently encouraging the section off the blade, and place them into a flotation bath, laying the section flat.

13.   If rippling occurs, place the block back onto the cold plate for 5-10 minutes.

14.   To mount the section on a glass slide, dip the slide into the bath, make contact with the side of the section, and pull up out of the water bath.

15.   Label the slide immediately with your name, date, and tissue, and place it in a drying oven.

16.   To remove unwanted sections from the water bath, use a tissue to gather them by dragging it across the surface of the water.

17.   Clean all equipment well after use to ensure the area is ready to be used with no risk of contamination.