Saturday, September 12, 2009

cytogenetics[THERMOTRON & TECAN harvester]

Hello everybody,
in the blink of an eye, we are ending our attachment in less than 2months time. Hope everyone is still doing managing well. Anyway, back to the topic, i will be focusing about the two machines that are used in the cytogenetics lab. Cytogenetics is the study of chromosomes structures, its function and behaviour.

TECAN harvester






[The reagents]



As the name implies, it is a harvester however it is a robotic harvester that is involved in the harvesting of the cells. There are three major steps in the Harvesting procedure. The first step is Mitotic arrest,so as to arrest the cells in the metaphase stage because chromosomes are best studied at the metaphase stage as it is the clearest and at its most contracted state. The tecan harvester will add in Colcemid, a mitotic inhibitor that helps prevent spindle fiber formation, a process by which sister chromatids are pulled to opposite poles for incorporation into the 2 daughter cells. In addition,it also promotes chromosome condensation.

The second step is Hypotonic treatment whereby the cells are treated with a hypotonic saline solution to increase the cell volume so that chromosomes can spread out to allow easier identification.

The last step in harvesting is Fixation whereby the cells are fixed with addition of 3:1 methanol: glacial acetic acid so as to remove water from the cells and preserve them, by hardening the membrane and prepares chromosomes for the bending procedure.




[Drying of slides]

After which the slides are dried in the THERMOTRON before they are heated, stained and karyotyped.

hOPE this will give everybody some idea how harvesting of cells is carried out in the cytogentics lab that i am attaced to.

cheers,
Yong Herng
0702243G
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6 comments:

  1. emadtechsSeptember 13, 2009 at 5:51 AM

    Hey man !

    Hope you are well =)

    Wow, so cool you have a robotic harvester ! I had 31 samples to harvest last friday and i'm sure one in my lab will help !

    I'm curious to know more about your slide dryer. Is it like an incubator whereby you put the slides inside and the conditions of temperature and humidity are controlled ? If so, what are the temperature and humidity settings used ?

    Thanks man, hope you enjoy cytogenetics =)

    Take care !

    Ng Tze Yang Justin
    0703747F

    ReplyDelete
  2. Sesame Chicken (:September 13, 2009 at 6:47 AM

    Hey YH,

    Wont the Chromosome become denature when put to dry? DNA and chromosome materials are fragile stuff.

    At what temperature does the THERMOTRON dries the sample and for how long?

    Cheers
    Tiong Han
    Tg01

    ReplyDelete
  3. Sesame Chicken (:September 14, 2009 at 6:43 AM

    To Justin:
    Hey haha yea i heard the cytogenetics lab i am attached to is the only lab that uses the Tecan harvester. It certainly reduces time and human error with the robotiv harvester. Anyway, the Thermotron indeed has a controlled temperature and humidity setting where the temperature is 25 degree celsius while the humidity is set at 40%. This conditions is fixed and set for optimal drying.

    See you on fri!
    Cheers,
    Yong Herng


    To TH,
    the temperature for the thermotron is set at 25 degree celsius, at such a temperature below room temperature, the chromosomes would not denature so easily. There is no fixed time for the sample to dry, after sucking the media out of the small petri via a tube(As seen in picture), you leave the small petri dish containing the coverslip inside the Thermotron for around a few minutes. Once you see that the coverslip is clearly dry, you can bring them out.

    Cheerrs
    Yong Herng

    ReplyDelete
  4. TG01 Group 1September 15, 2009 at 4:00 AM

    HI there!
    the 3 steps are all done within the same day is it?

    thanks!

    stella
    0701059H

    ReplyDelete
  5. Sesame Chicken (:September 19, 2009 at 8:24 PM

    hey tere!
    yup, is all done in the same day, with the robotic harvester,the 3 steps are done in around half an hour.

    cheers!

    Yong Herng
    0702243G

    ReplyDelete
  6. chenOctober 22, 2009 at 5:33 PM

    Hi! May i know what exactly is the colcemid that used as mitotic inhibitor? Usually we use colchicine, isn't it?

    Regards,
    Kok Siong Chen
    http://www.treatgene.com

    ReplyDelete

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