Mentors at Your Benchside

#85 — Working with living cells is a tricky business, and tiny fluctuations in environmental conditions can affect their physiology and impact your experiments. Or worse, it can lead to their death. Game over!

Passaging your cells involves removing them from their growth medium to transfer them to fresh vessels with fresh media.

In this episode, learn how to passage adherent and suspension cells while minimizing the impact on them. See the different passaging strategies you can adopt and decide which ones are right for you!

Check out the corresponding online article for handy tables and figures. [1] Learn all about cell confluency and why it matters [2] and explore the critical components of cell culture media. [3]

Resources:
1. How To Passage Cells in Culture. Available at: https://bitesizebio.com/13680/how-to-passage-cells/
2. How Confluent Are Your Cells? A Beginner’s Guide to Measuring Cell Culture Confluency. Available at: https://bitesizebio.com/63887/cell-confluency/
3. What’s in Your Cell Culture Medium? Available at: https://bitesizebio.com/37034/cell-culture-medium/

#84 — Every experiment starts by preparing some buffer solutions. And every buffer solution starts with weighing out some compounds on an analytical balance.

But these essential yet sensitive pieces of lab equipment are prone to measurement drift—meaning you could be weighing out different amounts every time you use one.
 
In this episode, get three easy tips to avoid measurement drift and learn some reliability testing methods to check your lab balance and prepare solutions with confidence.

Check out the full online article here [1] and read our essential guide to cleaning and caring for lab balances. [2] Plus, check out these eight steps you can take to improve lab accuracy and precision. [3]

Resources:
1. 3 Easy Tips for Avoiding Measurement Drift in Analytical Balances. Available at: https://bitesizebio.com/33245/drift-measurements-analytical-balances/
2. How to Clean and Calibrate Your Lab Balance. Available at: https://bitesizebio.com/24193/how-to-clean-and-calibrate-your-lab-balance/
3. How to Measure and Improve Lab Accuracy and Precision. Available at: https://bitesizebio.com/55470/accuracy-and-precision/

#83 — Chemically competent cells are a key resource in molecular biology labs. But do you really understand what is meant by chemically competent?

In this episode of Mentors At Your Benchside, we explain the science behind them and share how you can make your own stocks, saving you money and avoiding nasty surprises when someone forgets to reorder stocks.

Be sure to visit the original article for helpful diagrams and tables, [1] download our free chemically competent cells cheat sheet for a handy print protocol, [2] and learn how to make your own electrocompetent cells. [3]

Resources:
1. How to Make Your Own Chemically Competent Cells. Available at: https://bitesizebio.com/30145/chemically-competent-cells/
2. Bitesize Bio's Chemically Competent Cells Protocol. Available at: https://bitesizebio.com/chemically-competent-cells-protocol/
3. DIY Electrocompetent E. coli. Available at: https://bitesizebio.com/969/diy-electrocompetent-e-coli/

#82 — Did you know that most centrifuge accidents result from user error and improper centrifuge care? While proper balancing of samples is important, it is not the only thing you need to be aware of when using your centrifuge. In this episode of Mentors At Your Benchside, we give you 5 tips to help ensure your centrifuge keeps spinning.

Read the full article to get the formula for finding the real maximum speed to use based on a solution’s density. [1] Need to use an ultracentrifuge? These beasts should be respected, but there is no need to be petrified of them; just read our tips on using an ultracentrifuge safely. [2] Finally, for more advice on using centrifuges, be sure to watch the on-demand webinar from Eppendorf on Essentials in Centrifugation to get informed on how centrifuges work, practical tips and tricks, and critical safety information. [3]

Resources:
1. How to Keep Your Centrifuge Alive: 5 Easy Tips. Available at: https://bitesizebio.com/31550/keep-centrifuge-alive/
2. Respect the Ultra. Available at: https://bitesizebio.com/4251/respect-the-ultra/
3. Essentials in Centrifugation - Better Safe than Sorry! (Webinar). Available on-demand: https://events.bitesizebio.com/essentials-in-centrifugation-1/join

#81 — You’re probably aware of the two main types of restriction cloning (sticky-end and blunt-end cloning), but do you know the difference? And do you know how to do both?

In this episode, we share what blunt-end cloning is, how it works, and give you tips and tricks for performing blunt-end cloning.

Visit the original article for detailed diagrams and helpful tables, [1] brush up on how DNA ligase works, [2] and reacquaint yourself with the different DNA polymerases. [3] You can also get help to improve your blunt end ligations, [4] and discover the magic of TA cloning. [5]

Resources:
1. A Beginner’s Guide to How Blunt-End Cloning Works. Available at: https://bitesizebio.com/45131/a-beginners-guide-to-how-blunt-end-cloning-works/
2. DNA Ligation: How it Works & 6 Top Tips.  Available at: https://bitesizebio.com/10279/the-basics-how-does-dna-ligation-work/
3. Get To Know Your DNA Polymerases. Available at: https://bitesizebio.com/24551/get-to-know-your-dna-polymerases/
4. 10 ways to improve blunt-end ligations. Available at: https://bitesizebio.com/19149/7-ways-to-improve-blunt-end-ligations/
5. Ta-Da! The Magic of Taq and TA Cloning. Available at: https://bitesizebio.com/19709/ta-da-the-magic-of-taq-and-ta-cloning/

#80 — Dive into the fascinating world of histology as we explore the basics of Hematoxylin and Eosin (H&E) staining, a cornerstone technique in tissue study. [1]

Whether you're a budding biologist or just curious about cellular structures, this episode of Mentors At Your Benchside is your introductory guide to the history of H&E staining, its mechanism of action, and the structures it effectively stains. Discover how the positive charge of hematoxylin stains nucleic acids blue, and how the negatively charged eosin brings out the pink in proteins.

And, if you want to go beyond hematoxylin and eosin staining, why not download our free histological stains poster and brighten up your lab? It lists common stains, the color they stain tissues, and some useful notes! [2]

Resources:
1. A Beginner’s Guide to Hematoxylin and Eosin Staining. Available at: https://bitesizebio.com/13400/a-beginners-guide-to-haematoxylin-and-eosin-staining/
2. Bitesize Bio's Histological Stains Poster. Available at: https://bitesizebio.com/histological-stains-poster/

#79 — Are you struggling to keep your proteins "happy" and active for your experiments? In this episode of Mentors At Your Benchside, we dive into the five critical elements you need to design the ideal protein purification buffer: pH, the buffer system, salt concentration, reducing agents, and stabilizing additives. [1]

Learn how buffers work, [2] and how to navigate the delicate balance of these factors to prevent protein aggregation and keep your purified proteins soluble and active. [3] Tune in for a simplified guide to mastering your protein purification buffer.

Resources:
1. 5 Ingredients for the Perfect Protein Purification Buffer. Available at: https://bitesizebio.com/7893/how-to-design-the-perfect-protein-purification-buffer/
2. How Do Buffers Work? Available at: https://bitesizebio.com/8478/how-do-buffers-work/
3. The Definitive Guide to pH, pKa, and pI. Available at: https://bitesizebio.com/9425/what-is-ph-pka-pi/

#78 — Want to visualize if your proteins interact in live cells? FRET is the answer. In this Mentors at Your Benchside episode, we explain how FRET works, why it's great for studying protein–protein interactions, and why it is not actually named Fluorescence Resonance Energy Transfer.

Visit the original article to see helpful diagrams, [1] brush up on how fluorescent molecules work, [2] and read up on how to measure FRET for the different methods and practical tips for choosing FRET pairs. [3]

Resources:
1. How FRET Works: A Guide to Visualizing Protein Interactions. Available at: https://bitesizebio.com/23012/how-fret-works/
2. How Fluorescent Molecules Work. Available at: https://bitesizebio.com/32973/fluorescent-molecules/
3. How to Measure FRET. Available at: https://bitesizebio.com/23295/how-to-measure-fret/

#77  — Have you ever thought about accessibility in science? We don’t always present our science in ways that are accessible to everyone. Nor is lab-based science always accessible.

In this episode of Mentors At Your Benchside, we explore what accessibility is and highlight how we can all make science more accessible and inclusive.

Visit the original article for helpful resources and to revisit the key definitions. [1] Make your presentations more accessible using the color blindness simulator, [2] the accessible color pallets, [3] and the list of accessible fonts. [4]

Resources:
1. Accessibility in Science: How Can We Make Science More Accessible? Available at: https://bitesizebio.com/65863/accessibility-in-science/
2. Coblis —Color Blindness Simulator. Available at: https://www.color-blindness.com/coblis-color-blindness-simulator/
3. Coloring for Colorblindness: Color Palette. Available at: https://davidmathlogic.com/colorblind/#%23D81B60-%231E88E5-%23FFC107-%23004D40
4. My favorite free fonts for print disabilities. Available at: https://www.perkins.org/resource/my-eight-favorite-free-fonts-print-disabilities/

#76 — Do you know how the first cells were identified? Or who discovered them? What about why they are called cells? Discover the fascinating history in this enlightening Mentors At Your Benchside episode.

Visit the original article for a timeline of cell biology, [1] discover the most commonly used cell lines, [2] and find out why HeLa cells are surrounded by controversy. [3]  

Resources:
1. History of Cell Biology. Available at: https://bitesizebio.com/166/history-of-cell-biology/
2. Top 5 of the Most Commonly Used Cell Lines. Available at: https://bitesizebio.com/33473/top-5-of-the-most-commonly-used-cell-lines/
3. The Story Behind Your Cell Culture. Available at: https://bitesizebio.com/7647/the-story-behind-your-cell-culture/