Peripheral Blood Smear Staining (F)

Question 1

What are the purposes of PBS?

Answer 1

1) Examination of the circulating blood cells under the microscope
2) Correlation the released blood cell cts from automated machines and obtained blood cell cts from manual methods
3) Confirmation of the presence of abnormal blood cells

Question 2

What are the purposes of PBS staining?

Answer 2

1) To make the circulating blood cells more visible under the microscope
2) To make the morphology of normal cells and abnormal cells more distinct under the microscope

Question 3

What is the most common stain used for PBS staining?

Answer 3

Combination of Wright and Giemsa

Question 4

What are the methodologies of PBS staining?

Answer 4

1) Staining Jar Method or Dip Method

2) Staining Dish Method

Question 5

The Dip method is recommended for what?

Answer 5

It is reco for staining a single slide

Question 6

What is the procedure (or steps; or stains w/ corresponding time) for dip method?

Answer 6

1) Methanol: 30 secs
2) Eosin: 6 secs
3) Methylene blue: 4 secs
4) Distilled water: 45 secs

Question 7

What is the purpose (or role) of methanol?

Answer 7

Fixative

Question 8

What is the purpose (or role) of eosin?

Answer 8

Acidic dye

Question 9

What is the purpose (or role) of methylene blue?

Answer 9

Basic dye

Question 10

What is the purpose (or role) of distilled H2O?

Answer 10

Buffer

Question 11

The staining dish method is recommended for what?

Answer 11

It is reco for staining multiple slides

Question 12

What is the procedure (or steps; or stains w/ corresponding time) of staining dish method?

Answer 12

1) Methanol: 30 secs
2) Eosin: 6 secs
3) Methylene blue: 4 secs
4) Distilled water: 45 secs

Question 13

What is the purpose (or role) of methanol?

Answer 13

Fixative

Question 14

What is the purpose (or role) of eosin?

Answer 14

Acidic dye

Question 15

What is the purpose (or role) of methylene blue?

Answer 15

Basic dye

Question 16

What is the purpose (or role) of distilled H2O?

Answer 16

Buffer

Question 17

What are the 2 types of evaluation of a PBS?

Answer 17

1) Macroscopic evaluation of blood film

2) Microscopic evaluation of blood film

Question 18

What is the color appearance of PBS for macroscopic evaluation?

Answer 18

The film should appear pink to purple

Question 19

What is the color appearance of RBCs in microscopic evaluation of PBS?

Answer 19

RBCs should be orange to salmon pink

Question 20

What is the color appearance of the nuclei of WBCs in microscopic evaluation of PBS?

Answer 20

The nuclei of WBCs should be purple to blue

Question 21

What is the color of cytoplasm and granules of neutrophil in microscopic evaluation of PBS?

Answer 21

The cytoplasm of neutrophil should be pink to tan w/ violet of liliac granules

Question 22

What is the color appearance of granules (including its characteristic) of eosinophils?

Answer 22

Eosinophils should have bright orange refractile granules

Question 23

What is the color appearance of PLTs in microscopic evaluation of PBS?

Answer 23

PLTs should be dark pink to lavender in color

Question 24

What are the results of improper staining of PBS?

Answer 24

1) RBCs appear gray
2) RBCs are too pale or are red
3) WBCs are too dark
4) WBCs are barely visible
5) Eosinophil granules are gray

Question 25

What are the problems (in terms of excessive blue stain) and their corresponding resolutions?

Answer 25

1) Stain or buffer too alkaline -> change your buffer and check the pH before using
2) Inadequate rinsing -> follow the prescribed # of secs in rinsing
3) Prolonged staining -> follow the prescribed # of secs in staining
4) Thick smears -> lower the angle of spreader slide
5) Heparinized blood sx -> only use blood sxs mixed w/ EDTA

Question 26

What are the problems (in terms of excessive pink stain) and their corresponding resolutions?

Answer 26

1) Stain or buffer too acidic -> change your buffer and check the pH before using
2) Prolonged rinsing -> follow the prescribed # of secs in rinsing
3) Inadequate staining -> follow the prescribed # of secs in staining
4) Thin smears -> raise the angle of spreader slide
5) Inadequate buffering -> follow the prescribed amt of buffer

Question 27

What are the common PLT morphology findings?

Answer 27

1) Giant PLTs
2) Stress or Reticulated PLTs
3) Red cell fragments
4) PLT clumps
5) PLT satellitosis
6) PLT-neutrophil agglutination