Mouse embryo anatomy

Mouse embryo (E12.5) dissection was performed so as to isolate and culture embryonic liver (last image). Liver appears red as during foetal development is the major site of erythropoiesis (i.e. the generation of blood cells). Third image shows the various developing internal organs of the GI tract at this point in the development of the embryo (i.e. after the outgrowth of the epithelium from the gut tube and the establishment of the various organ 'buds'). Images were taken from an iPhone 4 via a Leica stereomicroscope.

Peripheral Blood Film II: microscopy

Using light microscopy the morphology, colouring and consistency of red blood cells (RBC), white blood cells (WBC) and platelets can be determined. Various diseases associated with abnormalities in blood cells, such as haemoglobinopathies, anaemias, leukaemias etc., can be initially diagnosed from the film, however, further biochemical analysis is required.

The first image was taken at 400x magnification showing that the vast majority of cells found in peripheral blood are red blood cells, with only a few (3 in this image) white blood cells present. Further magnification at 1000x was used in the remaining images. There are several types of WBCs present in peripheral blood and tissues, with each type having its own distinct characteristics (morphological) that makes them different from the other types.

The images show the larger size of WBCs compared to RBCs and the presence of the nucleus (big purple object in the middle of the cell) which is absent in RBCs. The morphology of the nucleus, as well as the colour of the cytoplasm, can help in determining the type of WBC. In the second image, 3 WBCs are present. The one on the left being a polymorphonuclear (no fixed structure of the nucleus) eosinophil, as the cytoplasm contains several vesicles containing cytotoxic chemicals (granules) that are coloured slightly red from the dye and the nucleus has a non-fixed shape. The cell on the top right has a uniform big circular nucleus and low cytoplasm/nucleus ratio, suggesting that this cell is a lymphocyte. The right bottom cell has a nucleus with irregular shape and contains some vesicles/granules with the cytoplasm being almost colourless, suggesting that the cell is a polymorphonuclear neutrophil, the most abundant WBC in peripheral blood. The small purple dots that appear in the image in-between cells are platelets (fragments of cells that help in the coagulation process). The 3rd image shows a lymphocyte and a neutrophil and the last image 2 neutrophils, all surround by RBCs and platelets.

Peripheral Blood Film

After venipuncture (blood collection process from patient using a syringe) the peripheral blood is collected. A drop of blood on a microscope slide can be spread evenly using a cover slip so as to create an even smear (film). The spread has to be extremely thin at the very end, so as to create a single cell layer which will allow bloods cells to be viewed under the microscope.

After the spreading on the microscope slide, the cells on the slides can be stained using various techniques, such as the Giemsa stain.

Melathron Agoniston EOKA Biomedical laboratory

During the summer (Jun-Sep), I was recruited as a Biomedical laboratory Intern in Melathron Agoniston EOKA. It was an enjoyable and fascinating experience from which I gained a lot of additional scientific knowledge and practical skills, as well as developed many interpersonal abilities.

The pictures show the main area of the laboratory, in which all the specific biomedical tests were carried out. Moreover, there are 2 additional rooms which are part of the laboratory, one of them being used as an office of the Head of the Department (Dr. Ellina-Papaphoti Georgia M.D.) and the other one used for venipuncture.

Mouse (mus musculus) intestine

Cross-section of the small intestine of mouse (mus musculus) close to the duodenum. The lumen of the gut is at the top of each image. The first and last images were taken at 100x while the second and third showing villi of the epithelium lining the gut at 400x. Purple dots in the middle of each cell are the nucleus of each epithelial cell and pink is the cytoplasm of each cell.
The slide was prepared at the lab of the university, as part of a practical session, but the pictures were taken using my personal light microscope at home with a regular camera.

Onion Skin

Onion skin cells alignment at 100x.
Pictures were taken with my digital camera using my personal light microscope at home.

Immuno-histochemistry and In-situ Hybridization

Both pictures show mouse (mus musculus) embryos 9.5days old and 11.5d respectively.
In the top picture the embryo was hybridized using the anti-sense probe for FGF8 and detected by alkaline phosphatase conjugated antibodies which stained the specific sites of expression of FGF8. The areas in which the staining is more intense are the midbrain hindbrain boundary, forelimb bud and hindlimb bud, especially the edge of the hindlimb bud, the apical ectodermal ridge.
In the second picturethe embryo that was incubated with both primary antibodies and secondary conjugated with peroxidase antibodies, showing the specific sites of expression of neurofilaments. The visible staining represents neurofilaments, spinal ganglia (SpG) developing in the developing vertebrae and the developing cranial nerves, the vagus neurofilament, the glossopharyngeal (GlPh) neurofilament and the hypoglossal neurofilament.

Both pictures were taken from a camera connected to a dissection microscope.

Immunofluorescence

The green fluorescence represents the location in the cell of a protein found in the Golgi apparatus in all pictures.
The red fluorescence:
1) in the top picture represents the location of the cytoskeleton of the cell and specifically microtubules (anti-β-tubulin used)
2) Middle picture represents the location of late endosomes and lysosomes in the cell (anti-CD63 used)
3) in the bottom picture represents the location of the endoplasmic reticulum (E.R.) in the cell (anti-PDI used).

The cells used were COS-7 (African green monkey SV40 transformed kidney fibroblasts) and confocal microscopy was used to view the fluorescence emitted by the fluorophore of different color conjugated to secondary antibodies.

Haemoglobin Studies

Haemoglobin from horse red blood cells was purified, some was left to crystallize using different salt concentrations (top picture) and a sample was loaded on a polyacrylamide gel along with molecular weight markers. Sodium Dodacyl Sulfate - Polyacrylamide Gel Electrophoresis (SDS-PAGE) is a very useful technique for all bio-science researchers, giving relatively a lot of information about a protein (especially the 2-D gel).

Polytene Chromosome

Polytene chromosomes from the salivary glands of fruit fly larva (Drosophila Melanogaster) stained with lacto-acetic orcein and viewed under a light microscope at 400x.