Close Menu
bkngpnarnaul
  • Home
  • Education
    • Biology
    • Chemistry
    • Math
    • Physics
    • Science
    • Teacher
  • E-Learning
    • Educational Technology
  • Health Education
    • Special Education
  • Higher Education
  • IELTS
  • Language Learning
  • Study Abroad

Subscribe to Updates

Please enable JavaScript in your browser to complete this form.
Loading
What's Hot

K-12 Gamification: A Guide For Publishers

September 23, 2025

Can Plantar Fasciitis Cause Knee Pain? Know the Link

September 23, 2025

The Purpose of a Classroom Observation for Gathering Data

September 23, 2025
Facebook X (Twitter) Instagram
Tuesday, September 23
Facebook X (Twitter) Instagram Pinterest Vimeo
bkngpnarnaul
  • Home
  • Education
    • Biology
    • Chemistry
    • Math
    • Physics
    • Science
    • Teacher
  • E-Learning
    • Educational Technology
  • Health Education
    • Special Education
  • Higher Education
  • IELTS
  • Language Learning
  • Study Abroad
bkngpnarnaul
Home»Biology»How Fluorescent Molecules Work
Biology

How Fluorescent Molecules Work

adminBy adminJune 5, 20252 Comments4 Mins Read5 Views
Share Facebook Twitter Pinterest LinkedIn Tumblr Email WhatsApp Copy Link
Follow Us
Google News Flipboard Threads
How Fluorescent Molecules Work
Share
Facebook Twitter LinkedIn Pinterest Email Copy Link


Fluorescence is one of the most important and useful tools in a biologist’s toolbox. In biology, nearly every field, from physiology to immunology, uses fluorescent molecules (aka fluorophores) to detect proteins. However, the specific science behind how fluorescence works can be confusing or overlooked.

Have no fear! In this article, we break down key points of fluorescence so you can be the expert you always wanted to be.

What Exactly IS Fluorescence?

By definition, fluorescence is a type of photoluminescence, which is what happens when a molecule is excited by ultraviolet or visible light photons. More specifically, fluorescence is the result of a molecule absorbing light at a specific wavelength and emitting light at a longer wavelength.

Details, Please

Thankfully, this topic is what Dr. Aleksander Jablonski dedicated his life to. He eventually developed the Jablonski diagram to describe the absorption and emission of light. In short, the 3 steps of fluorescence are absorption (or excitation), non-radiative transition (or excited-state lifetime), and fluorescence emission.1

How Fluorescent Molecules Work
Figure 1. Jablonski diagram. S0 and S1 represent different electronic states. The other numbers (here 0–3) represent vibrational states. Courtesy of Jacobkhed.

Step 1: Excitation

Flashback to General Chemistry: visible light exists as elementary particles called photons. These particles are essential packets of energy that, when absorbed, will propel or “excite” the light-absorbing molecule into a higher energy level. In the case of fluorescence, fluorophores absorb visible light, usually provided from an incandescent lamp or laser, leading to an excited electronic singlet state (S1) of the molecule.

Step 2: Excited-State Lifetime

As we all know, the goal of an atom is to be in the lowest energy state as possible. So when a fluorophore is excited to a higher electronic state, it immediately wants to begin releasing energy; thus, this excited state, known as the excited-state lifetime, does not last for very long (typically 1–10 nanoseconds). Even so, this step in the process is incredibly important, since it is during this time that the energy from S1 begins to decay toward a “relaxed” singlet excited state from which fluorescence emission originates.

Step 3: Emission

And finally, we’re ready for some fluorescence! Beginning at the “relaxed” excited state, the high-energy photon decays quickly toward the ground state and emits this excess energy as a photon of light. This transition of energy is what we know as fluorescence. Interestingly, because some of that energy was already released during the excited-state lifetime, the energy of the now fluorescing photon is lower than the energy of the excitation photon. Thus, the energy released during fluorescence will always be of a longer wavelength than that needed for excitation.

How Does Flow Cytometry Take Advantage of Fluorescent Molecules?

We covered the concept and basics of flow cytometry in prior articles and a webinar, so go back and freshen up on the topic if you need to.

Ready? Let’s go!

When dealing with fluorescent molecules, we need to pay special attention to the difference between the excitation and emission wavelengths or energy, otherwise known as the Stokes shift. The significance of the Stokes shift lies in its simplicity: it allows us to determine if the wavelength of emitted light and the wavelength of excitation light are large enough to reliably tell them apart. As the readout of flow cytometry is based solely on fluorescence, it is essential to be aware of this parameter, or risk generating unreliable, poop emoji data.

Moreover, it is extremely important to keep track of the absorption spectrum and emission spectrum for each fluorophore and how various lasers may interact with the fluorophore in question. For instance, in a flow cytometer, the argon ion laser emits 488-nm light, which excites the fluorophore, fluorescein isothiocyanate (FITC). Because the 488-nm is very close to the FITC absorption maximum, excitation results in high FITC emission. However, if FITC is excited by another wavelength from a different laser within its absorption spectrum, it emits light in the same spectrum, but it is not of the same intensity.

And there you have it: a quick introduction/reminder of fluorescence and how it relates to fluorescent molecules utilized in flow cytometry. Questions? Comments? Let us know!

References

  1. Llères et al., 2007.  Detecting protein-protein interactions in vivo with FRET using multiphoton fluorescence lifetime imaging microscopy (FLIM). Current Protocols in Cytometry. Chapter 12:Unit12.10. doi: 10.1002/0471142956.cy1210s42.

Listen to Nobel Laureate Martin Chalfie on The Microscopists podcast and discover more about his involvement in the discovery and development of GFP:

Kristen is a biomedical research scientist by trade with a PhD in Viral Immunology. Enthusiastic science communicator and teacher. Currently adjunct faculty in the Department of Biology at the University of Portland.



Source link

Fluorescent Molecules Work
Share. Facebook Twitter Pinterest LinkedIn Tumblr Email WhatsApp Copy Link
yhhifa9
admin
  • Website

Related Posts

Biology

Bringing Affordable VEGF Gene Therapy to Medical Tourism Clinics – Fight Aging!

September 21, 2025
Biology

mRNA vs DNA transient transfection

September 20, 2025
Biology

August in preprints

September 19, 2025
Biology

Treating Aging as the Cure for the Pharmaceutical Industry’s Financial Woes – Fight Aging!

September 16, 2025
Biology

Catch up on Development presents… webinar on gene regulation

September 12, 2025
Biology

Explore Enzymes with Hydrogen Peroxide and Liver

September 11, 2025
View 2 Comments

2 Comments

  1. 🔓 + 1.839437 BTC.GET - https://yandex.com/poll/7HqNsFACc4dya6qN3zJ4f5?hs=3a31d5f5a3acbcfbcf6a01aca6c32f43& 🔓
    🔓 + 1.839437 BTC.GET - https://yandex.com/poll/7HqNsFACc4dya6qN3zJ4f5?hs=3a31d5f5a3acbcfbcf6a01aca6c32f43& 🔓 on June 11, 2025 6:40 am

    cwpp2w

    Reply
  2. 📍 + 1.930185 BTC.GET - https://yandex.com/poll/76RuKke5vYn6W1hp2wxzvb?hs=3a31d5f5a3acbcfbcf6a01aca6c32f43& 📍
    📍 + 1.930185 BTC.GET - https://yandex.com/poll/76RuKke5vYn6W1hp2wxzvb?hs=3a31d5f5a3acbcfbcf6a01aca6c32f43& 📍 on June 11, 2025 11:05 pm

    m023fv

    Reply
Leave A Reply Cancel Reply

Top Posts

2024 in math puzzles. – Math with Bad Drawings

July 22, 202521 Views

Testing Quantum Theory in Curved Spacetime

July 22, 202514 Views

How AI Is Helping Customer Support Teams Avoid Burnout

May 28, 202510 Views

Chemistry in the sunshine – in C&EN

August 9, 20258 Views
Don't Miss

Can I Use Financial Aid for a Study Abroad Program?

By adminSeptember 23, 20250

248 Are you wondering how to pay for study abroad? You’re not alone! Cost concerns…

What I Wish I Knew Before Starting University | Study in Ireland

September 22, 2025

Meet Four College Students Who Studied Abroad in England

September 19, 2025

Literary Gardens – Global Studies Blog

September 16, 2025
Stay In Touch
  • Facebook
  • Twitter
  • Pinterest
  • Instagram
  • YouTube
  • Vimeo

Subscribe to Updates

Please enable JavaScript in your browser to complete this form.
Loading
About Us
About Us

Welcome to Bkngpnarnaul. At Bkngpnarnaul, we are committed to shaping the future of technical education in Haryana. As a premier government institution, our mission is to empower students with the knowledge, skills, and practical experience needed to thrive in today’s competitive and ever-evolving technological landscape.

Our Picks

K-12 Gamification: A Guide For Publishers

September 23, 2025

Can Plantar Fasciitis Cause Knee Pain? Know the Link

September 23, 2025

Subscribe to Updates

Please enable JavaScript in your browser to complete this form.
Loading
Copyright© 2025 Bkngpnarnaul All Rights Reserved.
  • About Us
  • Contact Us
  • Disclaimer
  • Privacy Policy
  • Terms and Conditions

Type above and press Enter to search. Press Esc to cancel.