Live and stained microscopy (H&E and gram staining) of bacteria in single culture from previously taken mouth swab from Rhys 4/11/2017 & use of new SEP heater for heat-fixing and drying slides

In my previous posts, I explained how the children and I took a swab of Rhys’ mouth and cultured it. Then, we took sample of one of bacteria that grew on the first culture to grow a second culture of a single organism. Today, I created a number of slides from this second Petri dish and then viewed them using 20x, 32x, 40x and 63x objective (equivalent when you add in the magnifying effect of the Bresser MikrOkular camera to up to 4300x magnification (see next post – 63x objective = 4300x magnification on screen) with live culture, H&E staining and Gram staining.

This represents my most successful observations of bacteria under a microscope so far.

The appearance and staining properties of the bacteria show that they are gram positive rods. Gram positive rods and filaments are organisms that are commonly isolated from the biofilms of dental plaque. The following organisms are commonly found:

  • Actinomycetes
  • Lactobacilli
  • Eubacteria
  • Propionibacteria

I am not sure which of these we have grown here!

Andy

Heat fixing slides for staining using new variable heater:

63x objective live sample (depending on size of the screen you are using to view this image this equates to around 4300x magnification), clearly showing strings of long bacteria:

Phase contrast (Ph I annulus) 63x objective:

H&E staining, 63x objective – with this stain purple represents nuclear material and cell membrane and pink the cytoplasm. As you can see, at this enormous magnification (4300x) sub-structural differentiation (i.e. clearly different parts – different areas are blue and pink) can be seen within the bacterial cells:

Gram stain, 63x objective – Gram positive bacteria are purple suggesting this is Gram positive bacteria:

Although the following image does not add any detail or information to those above, it does represent an increase in magnification. I used the Bresser Mikrocam 9.0 camera to photograph the H&E slide and then cropped a small section of the photograph and used the high definition in the image to allow me to magnify the image further. The above images did not involve any post-processing but the image below has been post-processed in GIMP2 using some sharpening, contrast enhancement and some enhancement of colour. As a result of this, the magnification has been increased on screen to roughly 8000x magnification!!:

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