78 MICROSCOPY
Fig. 4. Monitoring cell growth ensures a standardised process. Choosing the right time to process proliferating cells is vital, ensuring they have grown enough for a sufficient yield, but have not yet reached saturation
technology removes the need to individually change the ring slit when switching from 4X-40X objectives. Moreover, enhancing observation of stem cell colonies, a new contrast technique known as inversion contrast (IVC) has been developed by Olympus. Tis novel method extends phase contrast to generate clear, artefact-free images with enhanced 3D information, delivering a greater level of optical information, especially from thicker samples such as induced pluripotent stem (iPS) cell colonies.
The process of monitoring cell growth Cell culture growth undergoes three distinct phases (Fig. 3.), and it is crucial to prevent the culture from proliferating beyond the log phase, where growth slows. In practical terms, the culture is ready to passage when confluency lies between 70-80%, and estimations are often made visually, which can be highly variable. Advances in software technology have now introduced accuracy and standardisation to confluency measurements for adherent cells (Fig. 2.).
www.scientistlive.com
Trough employing such software with a cell culture microscope, quantifiable cell growth data is quickly generated to ensure cells are always passaged at the correct time (Fig. 4.). Moreover, this enables scientists to create an accurate growth log, avoiding unnecessary dissociation and in-solution counting for the optimisation of culture conditions, for example.
Processing cells Whether preparing cell samples for passaging, downstream experimentation or storage, accurate in-solution cell counts are vital:
l Passaging – consistent seeding densities mean cultures grow at the same rate and health
l Downstream experimentation – identical cell counts facilitate comparable results
l Storing – knowing the cell concentration and viability in each vial is important when reviving cell lines
Traditionally, this count is achieved manually with the
haemocytometer. However, this slow and laborious technique introduces variations due to human error. A modern alternative is automated cell counting systems, which create an accurate cell count report in just 15 seconds that can be stored and exported, for streamlining, standardising and documentation (Fig. 2.).
Summary Cell culture forms the cornerstone of life science applications, and high quality results demand a high quality cell cultivation process. Tis can be enabled through considering many parameters and adopting the latest laboratory technologies available. Scientists are supported in perfecting a fast workflow that is fully documented for future records, and highly standardised to achieve successful life science experiments and regenerative medicine applications.
For more information ✔ at
www.scientistlive.com/eurolab
Jan Barghaan is with Olympus.
www.olympus-lifescience.com
Page 1 |
Page 2 |
Page 3 |
Page 4 |
Page 5 |
Page 6 |
Page 7 |
Page 8 |
Page 9 |
Page 10 |
Page 11 |
Page 12 |
Page 13 |
Page 14 |
Page 15 |
Page 16 |
Page 17 |
Page 18 |
Page 19 |
Page 20 |
Page 21 |
Page 22 |
Page 23 |
Page 24 |
Page 25 |
Page 26 |
Page 27 |
Page 28 |
Page 29 |
Page 30 |
Page 31 |
Page 32 |
Page 33 |
Page 34 |
Page 35 |
Page 36 |
Page 37 |
Page 38 |
Page 39 |
Page 40 |
Page 41 |
Page 42 |
Page 43 |
Page 44 |
Page 45 |
Page 46 |
Page 47 |
Page 48 |
Page 49 |
Page 50 |
Page 51 |
Page 52 |
Page 53 |
Page 54 |
Page 55 |
Page 56 |
Page 57 |
Page 58 |
Page 59 |
Page 60 |
Page 61 |
Page 62 |
Page 63 |
Page 64 |
Page 65 |
Page 66 |
Page 67 |
Page 68 |
Page 69 |
Page 70 |
Page 71 |
Page 72 |
Page 73 |
Page 74 |
Page 75 |
Page 76 |
Page 77 |
Page 78 |
Page 79 |
Page 80 |
Page 81 |
Page 82 |
Page 83 |
Page 84