The Invasion Assay

Bhuvi M -

Hello everyone! This week, I have been working on a poster based on my original research in Trimester 1, which I will present later this week at the Junior Science and Humanities Symposium.

In this blog post, I will focus on the Invasion Assay conducted during Trimester 1, as it laid the foundation for my research thus far.

The invasion assay marked the inception of my experimental exploration, aiming to assess the invasive capabilities of various cell lines, including OE33 (the negative control), GD, and FLO-1 (the positive control). Operating within the framework of a 24-well Corning invasion assay plate, each well has an inner well containing Matrigel.

Encountering a notable challenge of cell detachment due to the amount of cell growth, I navigated through this hurdle by using a hemocytometer—a precise method of cell counting.

Creating an environment conducive to cell invasion required establishing a nutrient gradient. I placed serum-free media in the inserts with suspended cells above the Matrigel, while serum-enriched media occupied the wells below the inserts.

After the initial 24 hours, no invasion was observed in any cell line. However, a pivotal development unfolded after 48 hours when both the positive control and GD cell lines exhibited clear signs of invasion. This led us to a crucial conclusion—that the afatinib-resistant cell line demonstrated invasive properties, indicative of a mesenchymal state. Our anticipation of the expression of mesenchymal markers in this cell line proved accurate. In contrast, OE33 exhibited an inability to digest the Matrigel, implying a non-mesenchymal state.

In summary, the Invasion Assay offered valuable insights into the invasive characteristics of different cell lines. Despite initial challenges, the results illuminated the mesenchymal state of the afatinib-resistant cell line and its potential for mesenchymal marker expression, distinguishing it from the epithelial state of OE33.

Looking ahead, I plan to delve into more experiments conducted during Trimester 1 and elaborate on how I am validating my findings through new experiments.

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    Alana Rothschild
    Wow- your post is very detailed and thorough. Thank you! Have you used a hemocytometer before? Can you give a brief description of how it helps count cells? I appreciate you enlightening me with all your information!
    Thank you, Ms. Rothschild! We use the hemocytometer whenever we need to count cell concentration. A hemocytometer is a specialized laboratory tool used to count and measure the concentration of cells in a liquid sample. It consists of a microscope slide with a gridded chamber, facilitating accurate cell counting under a microscope. Researchers use hemocytometers to assess cell density, viability, and other parameters in various biological and medical applications. Many prefer it to a digital cytometer as they can visually see which cells have died and which ones are alive.

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