Medical Students Corner

Hub For Medical Students

  • Home
  • ALL MBBS Books
    • 1st Year MBBS Books:[Free PDF]
    • 2nd Year Mbbs books[Free PDF]
    • 3rd year mbbs Book[Free PDF]
    • Final Year mbbs Books[Free PDF]
  • All Medical/ MBBS/BDS Topics MCQS
  • Medical Books
    • Anatomy
      • Histology
      • Embryology
      • NeuroAnatomy
      • Anatomy MCQs Book
      • Anatomy coloring book
      • Biochemistry
      • Physiology
        • Physiology MCQs Book
    • Pharmacology
    • Pathology
    • Microbiology
    • Biotechnology
  • Medical Topics & Softwares
  • BOARD EXAM BOOKS
    • USMLE
    • OET
    • MRCS
    • PLAB
    • FCPS
    • MRCP
  • Medicine
    • Cardiology
    • Pediatrics
    • Community Medicine
    • Forensic Medicine
    • Gastroenterology
    • Nephrology
    • Neurology
    • PEADIATRICS
    • Psychiatry
    • Pyschology
    • Radiology
    • Dermatology
    • Forensic Medicine
  • Surgery
    • Cardiology Surgery
    • Gynecology
    • Obstetrics
    • NeuroSurgery
    • Orthopedics
    • PEADIATRICS Surgery
    • Ophthalmology
    • ENT
  • GENERAL BOOKS
    • NOVELS
    • Poetry
  • Diseases And Managment
  • Medical Topics
    • Anatomy Contents
    • Physiology Contents
    • Pharmacology contents
    • Cases
  • DENTAL/ BDS
  • Medical Videos Lectures
  • NURSING BOOK
    • NBME
    • NCLX-RN
    • NEET
  • Privacy policey
  • Contact

Blood Smear (blood film) Preparation Procedure & Staining:

February 21, 2017 by dramjad Leave a Comment

The purpose of preparation of blood smear (blood film} is to study the morphology of RBCs, differential leukocyte count and reticulocyte count. Blood smear & Staining

Table of Contents

Toggle
  • APPARATUS:
  • Leishman’s Stain:
    • PRINCIPLE:
    • PROCEDURE:
  • A) Preparation of Blood Smear
    •  A good smear has following characteristics:
    • B) Staining of Blood Smear
    • (C) Examination of Smear Under Oil Immersion Lens:

APPARATUS:

4-5 glass slides, compound microscope, pricking needle (blood lancet), spirit swab, cedar wood oil/liquid paraffin, Leishman’s stain, wash bottle, buffered water and staining tray.

Leishman’s Stain:

1.5 gm powder of Leishman’s stain is dissolved in one litre of acetone free methyl alcohol. Leishman’s stain contains two dyes, eosin and methylene blue. Eosin is an acidic dye that stains basic structures like RBC and granules of eosinophil. It is pink or red in colour. Methylene blue is a basic dye that stains acidic structures like nucleus or granules of basophils. It is blue in colour. Acetone free methyl alcohol is a fixative for smear.

  1.  Fixation of smear is because of precipitation of proteins by alcohol which prevent washing off of the film.
  2.  It preserves the cells in what ever chemical and metabolicstate they are at the time of staining.
  3.  Acetone if present will cause shrinkage or even lysis of the cells.

PRINCIPLE:

Blood smear is prepared, stained with Leishman’s stain and cells are identified under oil immersion lens.

PROCEDURE: Blood Smear & Staining

A) Preparation of Blood Smear

  1. Selection of a spreader: Take one slide a spreader which has smooth edge. It should be done by careful look on the narrow edge of the slide or by moving a thumb smoothly on its edge. But, the slide should be washed with soap and water after touching its edge, to remove grease particles from its edges.
  2. Take 3-4 clean and dry glass slides and keep them on filter paper or any clean white paper placed on the table.
  3.  Prick the ring finger of left hand with the help of prickling needle under all aseptic conditions (discard the first two drOps of blood from skin puncture site). Specimen of blood can also be taken by venepuncture. Put a small size of blood drop on each glass slide about half centimeter from it’s narrow edge on the right side. Place the blood drop In the same way on other slides also.
  4.   Now put the spreader on left side of blood drop at the angle 45° as shown in Figure 14.
  5. Give left to right movement to the spreader so that blood comes along the edge of the spreader. Further give side-to-side movement so that blood comes along the whole edge of spreader.
  6. Now spread the blood by giving smooth, uniform and rapid movement to the spreader upto the left edge of the slide. Repeat the same procedure with rest of the slides.
  7. Dry the smear by waving the slides in the air for some time.
  8. Now observe the all smears whether they are satisfactory or not ).

 A good smear has following characteristics:

  • It is tongue shaped having head, body and tail. Head is the area where blood drop is placed. Body is the area between head and tall. Tail is the last part of the smear with ragged margins.
  • It should cover two-thirds of the slide.
  •   It should not be thick.
  •  There should not be marks or blank spaces in the smear.
  •  The dried film should be of brownish yellow colour (Buff coloured).

9. After selecting a good smear with naked eyes focus it under low power of the microscope. The smear should be thin enough to have single layer of cells. There

should not be rouleaux formation or clumping of cells.

B) Staining of Blood Smear

  1.  Leave it for 1-2 minutes for fixation of the smear.
  2. Add Leisman’s stain drop by drop till it covers whole of the smear. Count the number of drops you have put.
  3. Place 3-4 good slides horizontally on the stand.
  4. Add equal number of drops of buffered water (pH 6.8) on the slide. Mix the stain with water by blowing air with the help of a glass tube or with a dropper.
  5. Wait for 8-10 minutes for staining to complete. During this period in the presence of buffered water staining is taking place because of formation of cations and anions of basic and acidic dyes respectively. Methyl alcohol is unable to ionise the stain so unable to stain the cells.
  6. Wash the smear in slow running tap water or with the help of wash bottle till the smear becomes pink in colour. Clean the back of the slide to remove the stain from back side.
  7. Let the slide dry and focus it under high power. If it is under-stained, stain it again and if it is over-stained wash it again.

8.The best stained film should be selected for the examination of the cellular details.

(C) Examination of Smear Under Oil Immersion Lens:

Focus the slide under high power. Put a drop of cedar wood oil or liquid paraffin on the

slide and Shift the Oil immersion lens by constantly looking from the side of the microscope so that it just dips in the oil.

Now focus with the help of fine adjustment screw and examine the various cells for their identification. (Identifying features of various cells are given in Table 9.1. The various features of different cells are also shown in Figure 16.

DISCLAIMER:

Here i need you attention to some of the important words about this article. This post was share just for educational purpose and to help medical students, post graduates and professors. This Site only educate our visitors about the materials. Medicscenters.com do not share the copyright materials on this site. The Link which is provide as found/ taken from the others websites. we highly encourage our visitors to buy the materials from its original authors. More over we do not store any kind of illegal link in our medicscenter.com server. We always follow the legal DMCA policy. There is no copyright materials share on medicscenter.com, We only offer the materials which are free on internet. This website is educational purpose and if there is any copyright materials against DMCA policy, who want to remove the materials kindly contact us on email [email protected].

Filed Under: Physiology Tagged With: preparation of blood film, Preparation of blood smear, principle of blood smear and staining, procedure of blood smear and staining, staining

Leave a Reply Cancel reply

Your email address will not be published. Required fields are marked *

Recent Posts

  • Textbook of Psychiatry 3rd Edition PDF Download: 2025
  • The Maudsley Deprescribing Guidelines PDF Download:
  • Ethics and Professional Issues in Couple and Family Therapy PDF Free Download:
  • Gabbard’s Treatments of Psychiatric Disorders 5th Edition PDF Free Download: 2025
  • Study Guide: Introductory Textbook of Psychiatry 7th Edition PDF Free Download: 2025

Recent Comments

  • Lippincott Physiology PDF Download LINK [2020 Edition]: on All Anatomy and Physiology Textbook PDF Free Download [2024 direct link]
  • lippincott Biochemistry 7TH EDITION PDF DOWNLOAD LINK: on BRS Biochemistry PDF 7th Edition DOWNLOAD:2023
  • Critical Care Nursing: Diagnosis and Management PDF 8th Edition [Direct Link] - Medical Students Corner on Davis’s Drug Guide For Nurses PDF 17th Edition Free Download:2023
  • https://www.cheats-network.com on Jaypee Gold Mini Standard Atlas Orthodontics PDF Free Download:
  • anu on Tara Pharmacology 5th Edition PDF Direct Download

Archives

  • May 2025
  • April 2025
  • March 2025
  • February 2025
  • January 2025
  • December 2024
  • November 2024
  • October 2024
  • September 2024
  • August 2024
  • July 2024
  • June 2024
  • April 2024
  • March 2024
  • February 2024
  • January 2024
  • December 2023
  • November 2023
  • October 2023
  • September 2023
  • August 2023
  • July 2023
  • June 2023
  • May 2023
  • April 2023
  • March 2023
  • February 2023
  • January 2023
  • December 2022
  • November 2022
  • October 2022
  • September 2022
  • August 2022
  • September 2021
  • March 2020
  • February 2020
  • September 2019
  • February 2018
  • May 2017
  • April 2017
  • March 2017
  • February 2017
  • August 2016

Categories

  • 1st Year MBBS Books:[Free PDF]
  • ALL MBBS Books
  • All Medical/ MBBS/BDS Topics MCQS
  • Anatomy
  • Anatomy coloring book
  • Anatomy content
  • Anatomy Contents
  • Biochemistry
  • Biotechnology
  • BOARD EXAM BOOKS
  • Cardiology
  • Cardiology Surgery
  • Community Medicine
  • DENTAL/ BDS
  • Dentistry Books
  • Dermatology
  • Diseases And Managment
  • Embryology
  • ENT
  • Entrance Examination Guidelines
  • Forensic Medicine
  • Forensic Medicine
  • Gastroenterology
  • GENERAL BOOKS
  • Gynecology
  • Histology
  • Medical Books
  • Medical Topics
  • Medical Topics & Softwares
  • Medical Videos Lectures
  • Medicine
  • Microbiology
  • MRCP
  • MRCS
  • NBME
  • NCLX-RN
  • NEET
  • Nephrology
  • NeuroAnatomy
  • Neurology
  • NeuroSurgery
  • NOVELS
  • Nursing
  • NURSING BOOK
  • Obstetrics
  • OET
  • Ophthalmology
  • Orthopedics
  • Pathology
  • PEADIATRICS
  • PEADIATRICS Surgery
  • Pediatrics
  • Pharmacology
  • Physiology
  • Physiology MCQs Book
  • PLAB
  • Poetry
  • Psychiatry
  • Pyschology
  • Radiology
  • Radiology
  • Results
  • Surgery
  • Surgery Content
  • Surgery Contents
  • Uncategorized
  • Urology
  • USMLE

Meta

  • Log in
  • Entries feed
  • Comments feed
  • WordPress.org

Copyright © 2025 · Magazine Pro Theme on Genesis Framework · WordPress · Log in