supravital staining

Question 1

are young RBCs which are formed when the nucleus of the late normoblasts are lost through ___
- derived from the fact that the cell contains a small network of
basophilic materials called ___ which is only demonstrable by the use of supravital stain.
The more mature the reticulocyte is, the lesser the __of the cell.
- The __ is retained

Answer 1

Reticulocytes
- extrusion
- reticulum
- ribosomal content
- cytoplasmic RNA

Question 2

Reticulocyte count is used as an index of __ and ___
> Can be used to classify ___
> Also used to monitor therapeutic measures for anemia
> The specimen of choice is __

Answer 2

bone marrow activity and RBC production
- anemia
EDTA-anticoagulated whole blood

Question 3

• insufficient erythropoiesis and ineffective erythropoiesis

Answer 3

Bone marrow is unresponsive

Question 4

increased erythropoietic activity = increased reticulocytes in the circulation

Answer 4

Bone marrow can respond

Question 5

3 common supravital stains

Answer 5

new methylene blue - commonly used/ sharp blue color
brilliant cresyl blue
crystal violet

Question 6

Based on the property of ___ to react with basic dyes such as ____ or ____ to form a ___ of granules or filaments

Answer 6

Principle oof supravital stain
- ribosomal rna
- new methylene blue or brilliant cresyl blue

Question 7

Procedure
1. Mix equal volumes of__ and __ in a tube. Allow this mixture to stand at __ for __
2. Remix the preparation and prepare smears. Allow the smears to dry
3.Examine under __
4. Count the reticulocytes seen in __ successive fields of vision while
enumerating ___.

Answer 7

DRY METHOD
- blood and freshly filtered stain
- room temperature for 15-30 mins
- OIO
- 10
- 1000 mature RBCs

Question 8

Mature RBCs stain ___ while reticulocytes are identified by the presence of___ or __. Any cell
that contains two or more particles of blue-stained materials is classified as ___

Answer 8

gray-blue
deep blue filamentous web
granules within the cells
reticulocyte

Question 9

% Reticulocyte =

Answer 9

(# of reticulocytes/1000) x 100
or
# of reticulocytes/10

Question 10

DRY METHOD Normal Values
Conventional:

Answer 10

Adults: 0.5% - 1.5%
Infants: 2.0% - 6.0%

Question 11

DRY METHOD Normal Values
SI:

Answer 11

Adults: 5-15 x 10^-3
Infants: 20-60 x 10^-3

Question 12

In specimens with low hematocrit, the percentage of reticulocytes may be__

Answer 12

falsely elevated

Question 13

Corrected Reticulocyte Count (%) =

Answer 13

(%Reticulocyte x Patient’s Hct(%)) / 45

Question 14

should have an elevated corrected reticulocyte count of 2% to 3% to compensate for mild anemia

Answer 14

Hematocrit of 35%

Question 15

count should increase to 3% to 5% to compensate for moderate anemia

Answer 15

Hematocrit of less than 25%

Question 16

Actual number of reticulocytes in 1L (SI Unit) or 1ul (Conventional unit)

Answer 16

ABSOLUTE RETICULOCYTE
COUNT

Question 17

ABSOLUTE RETICULOCYTE
COUNT Conventional Unit
- Absolute reticulocyte count (uL) =

Answer 17

Reference Range: 20x10^3 - 115x10^3/ul
- = (% reticulocyte x RBC count (SI M/cumm))/100

Question 19

ABSOLUTE RETICULOCYTE
COUNT SI Unit
- Absolute reticulocyte count (uL) =

Answer 19

Reference Range: 20x10^9/L - 115 x10^9/L
- (% reticulocyte x RBC count (CU x 10^12/L))/100

Question 20

To evaluate the erythropoietic activity to represent one day production
- Correction should be made for the presence of shift reticulocytes if
___ is reported in the RBC morphology
- Reticulocyte is____ when polychromasia is present

Answer 20

RETICULOCYTE PRODUCTION
INDEX (RPI)
- polychromasia
- falsely increased

Question 21

released earlier to compensate for anemia
- matures 2-3 days in the circulation

Answer 21

Shift reticulocytes

Question 22

matures 2-3 days in bone marrow, 1 day in circulation

Answer 22

Normal reticulocyte

Question 23

RETICULOCYTE PRODUCTION
INDEX (RPI) formula

Answer 23

HCT (%)
________
RPI= % Reticulocyte x 45
_____________________________
Maturation time

RPI= Corrected Reticulocyte count
________________________________
Maturation time

Question 24

RETICULOCYTE PRODUCTION
INDEX (RPI) ref interval

Answer 24

RPI Bone marrow response
>3 Adequate/Increased
2-3 Normal
<2 Inadequate

Question 25

Patient Hct values to correction factor matration time (days)

Answer 25

40-45% 1
35-39% 1.5
25-34% 2
15-24% 2.5
<15% 3

Question 26

SOURCES OF ERRORS AND
COMMENTS of supravital stain

Answer 26

1 Anemic and polycythemic
> Adjust accordingly the blood to dye proportion
2 If blood and stain are not mixed before the films are made
> Should be mixed before making a smear because reticulocytes has lower specific gravity than that of mature RBCs
3 Moisture, poor drying
> forms refractile bodies
4. RBC inclusions that are also stained by supravital stains