groups » Laboratories discussion group » MALARIA SLIDE PREPARATION


Hi Awal and Ken,

How are you?
I have a question: we did another training with team and we had the same problem as I told you last time. We did the slide and I prepare the staining with distilled water with 4% giemsa (in 50ml in total), then after the staining we wait for 1hour, then we wash gently, please see our slide attached. I don't understand why it's look like that. if you could help it would be good for us.
I would like to send you and ken the pictures of this slide.

Please tell me I hear that another training will be done in tanzania for microscopist, is it will be the same microscopist group or with anothers persons.

Best regards,
Josiane.

Reply

  • kenawuondo KEN AWUONDO 9 Jul 2014

    Hi Josiane,
    Thick film made from EDTA blood easy to be washed off.
    My suggestion is:
    1. Make sure your slide glass is greasy-free slide.
    2. Spread the blood evenly (for the vaccine trial is 10ul blood for a rectangle 1x2cm of thick smear). Spread evenly using the pippet tip when you made the thick film.
    3. Dry the thick film using Slide warmer 37'C for 30 minutes. This drying method is important because you are now using EDTA blood.
    I do have the same problem if I rely the drying method is using air dried.
    If you dont have slide warmer: Dry the slide overnight, then stain the following day. (But this take time too long).
    4. Stain 4% for 45-60minutes (according to your SOP). Wash the slide gently after staining. If you rinse using squizing bottle, dont point to the thick smear directly. Rinse the label instead, and let water passing the specimen.

    Hope this help.

    Have a good day.

    Awal

    UCSF, Hanoi Vietnam

  • kenawuondo KEN AWUONDO 9 Jul 2014


    Dear Josiane,
    I am sorry that you encountered some problems while training. What volume of blood did you use? Can you make 5% working Giemsa solution and compare! It appears like your thin film is peeling off and this usually happen when the slide is greasy (not thoroughly cleaned).
    Please try 3uL of blood, 5% working Giemsa and stain for 45 mins then was with Buffer pH 7.2 and compare your results.
    Please feel free to contact us anytime.
    Many thanks
    Ken

  • lovafaa Love Ankrah 9 Jul 2014

    Aside all the suggestions given by our bosses, i will also like to stressed that,
    1. the stain should not be pour out from the slide before washing, meaning;
    ensured that the slide with the stain remained on the staining rack and start washing by continuously pouring the distilled water at the frosted or labelled end of the slide and not where the stained smear is.
    2.make sure the stain is completely washed off before you put the slide on vertical position for drying.
    3. if all things been equal never used lysed sample, else you wont see anything on the slide.

    all the best.

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