I prepare wet mounts using 5 ul of stained, PFA-fixed bacterial suspensions per slide with a cover slip (22 x22 mm) placed on them. Despite allowing the slide to stand for 5 minutes, there is a low of flow in the mount, making it almost impossible to image the cells.
How do I immobilise the bacteria without disrupting their cellular integrity?
Hi Shashank, have you tried fixing the sample with -20oC 100% methanol?
What about polylysine coating? It is simple and works well with living bacteria...
Steingrimur, isn't methanol affecting cell integrity?
I agree with Pierre. You either need to have a coating, such as gelatin, that enables the bacteria to stick to your slide or coverslip, or you need to imbed the bacteria in a viscous or immobile mounting solution. Tech tips at the following links:
https://www.researchgate.net/post/The_best_way_to_mount_bacterial_cells_for_fluorescence_microscopy2
https://www.agilent.com/cs/library/slidepresentation/Public/Sample%20Preparation_Immobilization%20and%20Imaging%20Dave%20Allison_032708.pdf
- Badges
- Science topic
- Similar topics
- Biological Science
- Microbiology
- Microorganisms
- Bacteria