Sunday, October 25, 2009

Slide Making

Hi everyone, I’m Justin here. Sorry for my super late post. For the last post, I’ll write about slide making.

Slide making involves dropping fixed cell suspension onto slides. Slides are then air dried. The air drying is based on the principle that fixed cells are supported with a thin layer of fixative. When the fixative evaporates, the cell would be pressed down, resulting in the spreading of chromosomes. The rate of fixative evaporation has to be controlled. If evaportation is too fast, a cytoplasmic background will be present and spreading will be poor. If the fixative evaporates too slowly, the weakened cell membrane may lose its integrity, leading to chromosome loss. Thus, the slide making process is able to eventually influence chromosome spreading and morphology.

The main factors that affect the slide making process are temperature, humidity and airflow. Humidity relates to the amount of moisture in the air. If the air is dry, evaporation rate will be faster, while if the air is moist, evaporation rate will be slower. Environment temperature is taken into account together as warm air is able to hold more moisture. The airflow within the area that slides are made affects the speed of drying. These factors directly affect the drying process, thus being the most important factors in a successful slide.

The lab I’m in uses a de-humidifier and the air conditioner for temperature and humidity. Another lab has a slide drying chamber, in which temperature, humidity, as well as air flow can be controlled more easily. It is important to consistently get slides of appropriate quality.

That’s all. Take care. To SIP and MP !

Ng Tze Yang Justin
0703747F
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4 comments:

  1. jenashling-shamunaOctober 26, 2009 at 10:11 AM

    Hi. erm for the fixing of cell suspension, the cell suspension was mixed with the fixative the dropped onto the slide or the cell suspension was dropped onto the slide first then the fixative was dropped onto the slide to fix the cell suspension on slide?

    Thanks

    Jennifer

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  2. The Madtechs Gp09 - TG02October 28, 2009 at 6:24 PM

    Hi Justin!

    Just wondering what type of fixatives did you use?

    And also, as you only mentioned about chromosomes, is slide making only for the purpose of screening chromosomes? How about looking at other cells in the slide?

    Thks!!

    Cheers,
    Zhang'e
    0704086H
    TG02

    ReplyDelete
  3. AnonymousNovember 2, 2009 at 4:28 PM

    Hey Justin!!

    I would like to know that since your lab does not have a slide drying chamber to control the airflow, how does your lab cope with uncontrolled airflow around the workplace? Or is airflow not as important a factor as temperature and humidity?

    Thanks!!

    Renee
    TG02
    0703634F

    ReplyDelete
  4. emadtechsNovember 4, 2009 at 1:42 PM

    Hi everyone,

    Sorry for my super late reply. Today is day 99 of SIP ! =)

    Jennifer,

    The cell suspension is fixed already. The slide making step follows after the harvesting.

    Zhang'e,

    The fixative is modified carnoy's fixative, using 3:1 methanol- acetic acid. Yea, the purpose of the cytogeneic workflow for karyotyping is to see chromosomes, so as to spot any abnormalities. Dividing cells are arrested using colcemid. For interphase cells, interphase FISH can be used to detect the specific abnormality.

    Renee,

    Yea everything is important. The problem is, many things are uncontrollable. To add airflow, a fan or mannual air pump can be used. Also, people walking near the slide making area can affect the airflow. Haha.

    Hope everyone is well =) To SIP, To MP ! =)

    Ng Tze Yang Justin
    0703747F

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