Why my section falling (coming) off slide? | How to prevent section from falling off slide? |
Insufficient fixation or unfixed tissues tend to come off slides more easily. | Fix tissues longer to get completely fixation of tissues. |
Formaldehyde fixed frozen sections are more prone to falling off slides. | Try to dry slides for much longer time or use alternative fixation such as acetone or alcohol. |
Tissue sections tend to come off slides more often on regular slides (uncharged or uncoated). | Always use positively charged or coated slides for immunostaining. |
You may just have had a bad batch of slides. | Replace with a new batch of slides. |
Hand lotion or cream contaminating the waterbath. | Avoid hand lotion or cream contaminating waterbath by wearing gloves. |
The disposable blades have oil on them. | Clean disposable blades with xylene. |
Wrinkles presented in the sections during the initial mounting may contribute to problem for falling off sections. | Try to spread the sections out and mount the sections on slides with wrinkle free. |
Paraffin sections may not be dried completely before placing in the oven. | Allow paraffin sections to air dry at least 30 minutes before placing in the oven at 56 C overnight. |
Frozen sections may not be dried completely before fixation and immunostaining procedure. | Allow frozen sections to air dry for at least 30 minutes before fixation and then air dry for another 30 minutes before immunostaining. |
Antigen retrieval procedure can cause sections to come off slides, especially when EDTA (pH8.0) or Tris-EDTA (pH9.0) such high pH antigen retrieval solution is used. | Use low pH solution such as citrate buffer (pH6.0) antigen retrieval Solution to replace EDTA (pH8.0) or Tris-EDTA (pH9.0) retrieval solution if it is possible. |
Distilled water alone may make sections come off slides easy. | Always use buffer solution to wash or rinse slides. |
Bone (especially the cartilage) tends to fall off slides after HIER. | Try other alternative retrieval methods such as enzyme digestion. |
Antigen retrieval devices may be trouble. | Try to use waterbath or steamer in stead of microwave or pressure cooker. |
Plain water in waterbath for mounting paraffin sections. | Add some gelatin in waterbath for mounting paraffin sections. |
If you have tried everything above and the problem persists. | Try gelatin coated slides and it has been working fairly well. |