By What is cytospin stain?
The cytospin technique uses a high speed centrifuge to concentrate the cells on a slide in a uniform monolayer 6 mm in diameter. Allow the slides to dry in air for several minutes and then stain them with Wright-Giemsa stain. Cytospin slides may be placed in an automatic stainer or stained manually.
What stain is used to stain a cytospin slide?
Two types of stain methods were used in this study: Giemsa and methylene blue/eosin. For methylene blue/eosin staining, the slides were fixed by placing 2 drops of fixing solutions on the slides and air dried in a biological hood (2 min) or on bench (3–5 min).
What is a cytospin instrument?
The Cytospin centrifuge is a special purpose instrument designed to deposit cells evenly onto a glass slide. The instrument when used correctly produces a monolayer cell deposition in a defined area of the slide using centrifugal force.
What is cytospin smear?
A cytocentrifuge, sometimes referred to as a cytospin, is a specialized centrifuge used to concentrate cells in fluid specimens onto a microscope slide so that they can be stained and examined.
What is the principle of H&E staining?
The principle behind H & E stain is the chemical attraction between tissue and dye. Hematoxylin, a basic dye imparts blue-purple contrast on basophilic structures, primarily those containing nucleic acid moeties such as chromtatin, ribosomes and cytoplasmic regions rich in RNA.
What is a cytospin used for?
A cytospin is obtained by employing centrifugal force to isolate, concentrate and deposit a monolayer of cells from a dilute cell suspension onto a circular area on a slide. The objective is to keep cells intact enabling the morphology of the cells to be examined.
What is the principle of Giemsa staining?
PRINCIPLE: The “neutral” dyes combining the basic dye methylene blue and the acid dye eosin, give a wide color range when staining. The pH of the staining solution is critical and ideally should be adjusted for different fixatives.
How to prepare cells for microscopy using cytospin?
Label the glass microscope slides. Assemble the glass slides, filter cards, and sample chambers in the cytospin centrifuge, ensuring that the centrifuge is balanced. Pipette 100–200 μl of cell suspension into each of the sample chambers. Centrifuge at 1000 × g for 5 min.
How is the settling rate of a Cytospin cell determined?
The fluid from the suspension is absorbed onto filter paper while the centrifuge is spinning. This allows the cells to adhere to the slide in a monolayer format. The cell settling rate is determined by the rotational speed of the centrifuge and the size and density of the cells.
What kind of resuspension is used in cytospin?
The type of resuspension solution used is dependent on the cells. A good rule of thumb is to use whatever solution will keep the cells alive and healthy throughout the cytospin procedure. The best morphology and subsequent staining of the cytospin cells are generated from cells that are freshly harvested.
How does the centrifuge work in a Cytospin cell?
A cytocentrifuge spins cells at an angle, at low speeds, and accelerates and decelerates gradually. The fluid from the suspension is absorbed onto filter paper while the centrifuge is spinning. This allows the cells to adhere to the slide in a monolayer format.