What can we see on this photomicrograph? - (Aug/23/2013 )

Hello,
i cultured some cocci on an agar plate and stored this plate for approximately one week at 4°C. Then, i examined the cells using a light microscope. Where I applied the cells to the object slide, I could observe the following image at 40x magnification.

http://imageshack.us/a/img694/4337/7a48.jpg

I don't think that these are cells but debris. But what exactly does this debris consist of? How does it emerge and how to avoid it? Why the debris is at a different layer than my cells (heat fixation)?

In terms of debris reduction, would it be better to culture the bactoeria in liquid medium or not storing the agar plate at 4°C.

Thanks for you responses.

To observe the cells properly you need to use a 100x objective.  From this picture you can't say much.

40x objective or total magnification?

Really hard to say anything

40x objective, 400x total magnification.

Hello,

I did Gram stain again. One more time, at 400x, I could see the above mentioned image.

With 100x magnification I have these photos. The first row is from one isolate. The second row from another isolate. I think it is Gram positive Corynebacteria. Do you agree with this?

I wonder that the cells on the right image of the first isolates looks a little bits coccoid.

For isolate 1, there are big round spots (maybe part of the club-shaped Corynebacterium?) The space between the spots sometimes isn't stained properly.

This doen not apply to isolate 2.

Also, I have a very basic question to which I could not find any answer in papers or university lectures.

WHAT exactly does Safranin stain? I just found a reference that it stains the nucleus but for bacteria??

You mean in the Gram staining? Safranin is just a contrast/counterstaining added to stain those cells that wouldn't stain otherwise by the first step. In fact, the safranin didn't appear in Gram's original protocol

Hello,
The image is from Gram staining :)
yes that's what I read but do you know to which structures of the cell safranin binds?
What do you think about the genus of my bacteria?

Cells appear to be coryneform.