Microscopy and Cells Flashcards
Why does histology have value in diagnosis?
It has great value because it is definitive. It means we can be certain what something is and therefore about what is the best treatment.
Define Biopsy.
The removal of a small piece of tissue from the body for examination under a microscope.
What are the 6 types of Biopsy?
Smear - Cervix/ Buccal Cavity
Curettage (spoon with sharp edge) - Endometrium of Uterus
Needle - large bored and comes out with tissue inside (organs)
Direct Incision - skin/mouth/larynx
Endoscopic - pincers remove tissue (lung/intestine/bladder)
Trans-vascular - pincers through blood vessel (heart/liver)
Define a tissue.
A collection of cells which are specialised to perform a particular function.
What is a microtome?
It is an instrument used to cut tissue sections so that they can be examined under the microscope.
What is the most common stain and what does it stain?
Haemotoxylin and Eosin.
H stains acidic components (such as the nucleus) purple and E stains basic components (cytoplasmic proteins) pink.
Periodic Acid Schiff stains mucus and mucus secreting glands a prominent colour. What is this?
Magenta
Why do biopsies need to be fixed?
A fresh biopsy is very wet and enzymes are present which will break down the structures. Glutaraldhyde/ Formaldehyde can be used to link macromolecules and so this preserves cellular structure and also kills any bacteria present.
Outline the basics of tissue processing.
Tissues need to be embedded in wax to be sectioned.
To get to this point they undergo
Dehydration - cleaning with ethanol - xylene/ toluene (wax adhesive)
Then: the reverse happens so it can be stained because most stains are water soluble.
Then: process happens again so that section does not degrade.
Why can tissue processing lead to shrinkage artefacts?
When tissue undergoes processing it goes through a number of chemical process and this can cause different parts of the sample to behave in different ways which can lead to changes of size - shrinkage artefacts
What is phase contrast microscopy?
This uses the interference effects when two waves combine to produce clearer images. This can be used to detail living cells.
What is dark field microscopy?
This is where an image is created from light scattering patterns.
How does fluorescent microscopy work?
It uses 3 stains which are fluorescent and can be used to highlight different parts of the cell depending on which antibodies they are coupled to.
What is the benefit of confocal microscopy?
It eliminates out of focus flare. It scans specimens with a laser beam and is able to produce 2D images which can then be built up into a 3D image. This process can be used on living specimens.
What is the Limit of Resolution?
Minimum distance at which two objects can be distinguished.