“Feasibility of Blue Ternate (Clitoria ternatea) Extract as an Alternative Supravital Stain for
Reticulocyte Examination in Peripheral Blood Smears”
Chapter 1
Introduction
Statement of the problem
1. What is the staining potential of Blue Ternate (Clitoria ternatea) extract in peripheral blood
smears?
2. How effective is Blue Ternate (Clitoria ternatea) extract compared to Methylene Blue in
staining blood cells?
3. Is there a significant difference between Blue Ternate (Clitoria ternatea) extract and
Methylene Blue in terms of clarity and visibility of blood cell structures?
4. What is the extent of staining quality provided by Blue Ternate (Clitoria ternatea) extract in
peripheral blood smears?
Statement of the Hypothesis
(Ho): There is no significant difference between the staining ability of Blue Ternate (Clitoria
ternatea) extract and Methylene Blue in reticulocyte examination of peripheral blood smears.
(H₁): There is a significant difference between the staining ability of Blue Ternate (Clitoria
ternatea) extract and Methylene Blue in reticulocyte examination of peripheral blood smears.
Significance of the Study
● This study looks at the use of Blue Ternate (Clitoria ternatea) flower extract as an
alternative supravital stain for reticulocyte examination in peripheral blood smears. The
results of this research are meaningful to several groups:
● For medical technology students, this study can serve as a learning tool to explore
natural and eco-friendly alternatives in staining procedures, which may also lower costs
for laboratory practices.
● For medical laboratories, the findings may suggest a possible substitute for Methylene
Blue, especially when standard stains are unavailable or expensive.
● For educators and schools, the study can provide an additional teaching reference that
shows how local plant resources can be used in basic laboratory work.
● For future researchers, this paper can serve as baseline information for further studies
on natural stains, helping improve methods and explore other plants as potential
alternatives.
Scope and Delimitation
● This study focuses on testing the feasibility of Blue Ternate (Clitoria ternatea) extract as
a supravital stain in reticulocyte examination. The research includes preparation of the
extract, blood collection using EDTA tubes, preparation of peripheral blood smears,
application of Blue Ternate stain, and microscopic observation of reticulocytes compared
with Methylene Blue.
● The study is limited only to the use of Blue Ternate extract in supravital staining. Other
staining techniques such as Wright’s or Giemsa are not included. The extract used is in
crude form and is not chemically purified. The study does not include clinical validation in
hospitals and is conducted only in a school laboratory setting.
● Despite these limits, the study aims to provide basic data on the potential of Blue Ternate
as an alternative stain, which may serve as a foundation for more advanced and wider
research.
Definition of Terms
1. Blue Ternate (Clitoria ternatea) Extract – A natural dye obtained from the flowers of the Blue
Ternate plant, used in this study as a possible substitute for Methylene Blue in staining blood
cells.
2. Supravital Stain – A type of stain applied to living cells to observe structures like RNA without
killing the cells.
3. Reticulocytes – Immature red blood cells that are newly released from the bone marrow into
the bloodstream.
4. Peripheral Blood Smear – A thin layer of blood spread on a glass slide for microscopic
examination of blood cells.
5. Methylene Blue – A standard laboratory supravital stain used to identify reticulocytes in blood
smears.
6. Staining Quality – The ability of a stain to make cell structures clear, visible, and
distinguishable under a microscope.
7. Feasibility – The possibility of using Blue Ternate extract effectively as an alternative
supravital stain based on clarity, visibility, and ease of use.
RRL& RRS
Foreign Study
1. Natural Dye from Butterfly Pea (Blue Ternate) in Blood Smear Staining
A study tested the aqueous crude extract from butterfly pea (Clitoria ternatea) flowers for animal
blood smear staining. Researchers found the extract could distinguish different blood
cells—they noted faint staining of nuclei, grayish-pink cytoplasm in red blood cells, and subtle
acidophilic (eosin-loving) staining in white blood cell granules. However, they also observed that
the stain's intensity was weak and recommended improving the extraction method and staining
conditions for better results .
https://www.researchgate.net/publication/385237621_EFFECT_OF_USING_AQUEOUS_CRUD
E_EXTRACT_FROM_BUTTERFLY_PEA_FLOWERS_CLITORIA_TERNATEA_L_AS_A_DYE_
ON_ANIMAL_BLOOD_SMEAR_STAINING?utm_source=chatgpt.com
Foreign Study
2. Blue Ternate Extract vs. Methylene Blue in Cheek Cell Staining
In a study published in 2023, C. ternatea flower extract was used to stain cheek (buccal) cells
as a safer, eco-friendly alternative to methylene blue. The findings were encouraging: the stain
produced clear sharpness and contrast, intensely stained cytoplasm evenly, and showed good
affinity for both the cell membrane and nucleus .
https://innspub.net/potential-of-clitoria-ternatea-l-flower-extract-as-a-safe-and-effective-alternativ
e-to-methylene-blue-stain/?utm_source=chatgpt.com
Local Study
3. Blue Ternate Extract as Primary Stain in Giemsa Method
Another study focused on using Blue Ternate flower crude extract as a primary stain for the
Giemsa method in blood smear analysis. They assessed key properties:
pH & solubility: The extract was mildly acidic (pH 5.2) and water-soluble, unlike Giemsa (pH 7.5,
also soluble) .
https://www.scribd.com/document/740682452/Blue-Ternate-Clitoria-ternatea-Flowers-Crude-Extr
act-as-an-Alternative-Primary-Stain-in-Giemsa-Method-in-Determining-Blood-Cells?utm_source
=chatgpt.com
Color intensity: Out of 10 slides, 60% stained equally intense compared to Giemsa, and 40%
stained darker. There was a statistically significant difference in color intensity (F = 6.0, p =
0.025) .
Visibility of blood cells: All slides stained with Blue Ternate defined blood cells, but uptake was
incomplete—white blood cells were not clearly visible. In contrast, Giemsa showed complete
uptake with clear cell visibility .
