Analytical and Quantitative Light Microscopy Course

mbl.eduAnalytical and Quantitative Light Microscopy | Marine Biological LaboratoryA comprehensive and intensive course in light microscopy for researchers in biology, medicine, and material sciences.

We are now accepting applications for Analytical and Quantitative Light Microscopy (AQLM) at the #MBL. Come see what you've been missing! www.mbl.edu/education/ad... #AQLM 🧫 #Microscopy 🔬 #WoodsHole 🦞 #Optics 💡 #LightMicroscopy 🧪 #FluorescenceMicroscopy ⭐️ #Confocal 🥼 #SuperResolution 🔥

Analytical and Quantitative Light Microscopy Course

Analytical and Quantitative Light Microscopy Course

The Speck Auditorium at the #MBL is a central gathering space for talks that bring together students, researchers, and visiting scientists from around the world. #AQLM 🧫 #Microscopy 🔬 #WoodsHole 🦞 #Optics 💡 #LightMicroscopy 🧪 #FluorescenceMicroscopy ⭐️ #Confocal 🥼 #SuperResolution 🔥

Lessons in #microscopy from Wendy Salmon, ALM: Tip 1: Patience makes it go faster. Take 0.5 seconds to make sure you won't crash the slide. Titrate that reagent to determine optimal staining. 📸1: #AQLM students learning single particle tracking with quantum dots with #OxfordInstruments. 📸2: Wendy!

Analytical and Quantitative Light Microscopy Course

Analytical and Quantitative Light Microscopy Course

Tip 5 continued: To define assay variables you want known samples (such as beads to define chromatic aberration if you're doing diffraction limited, multi channel localization comparisons). Known sample = an assay control #experiment. Does your assay do what you want it to?

Analytical and Quantitative Light Microscopy Course

Tip 6: The #scientific method states you should have ONE variable in your experiment. That's impossible in #biology, but be kind to yourself and tackle the assay variables separate from the biological variables, especially during early stages. #ScopeTheScope 🔬 #MBL 🌊 #AQLM 🧫 #Microscopy 🔬 #STEM🧬

Analytical and Quantitative Light Microscopy Course

Analytical and Quantitative Light Microscopy Course

Tip 4: Every experiment has a physical choreography to it. If what you want to do is hard or complicated, plan to practice (maybe even without a real sample). Even if everything else goes wrong with a first experiment, you've started figuring out the choreography.

Analytical and Quantitative Light Microscopy Course

Tip 5: What parameters you need for steps in a protocol are their own variables (staining concentration, type of #microscope, exposure time, averaging...). These are "assay variables" to distinguish them from the #biological question you're wanting to answer.

Analytical and Quantitative Light Microscopy Course

Analytical and Quantitative Light Microscopy Course

Tip 2: Maximize your likelihood for useful information. Do the #experiment to determine #antibody concentration or exposure time. Tip 3: The endeavor of #science is *figuring out how* to answer the question. Failure can't be completely avoided, though see Tip 1 to avoid total chaos.

Analytical and Quantitative Light Microscopy Course

#ThrowbackThursday with #AQLM! Registration for #AQLM2027 will open in the coming months. Stay tuned for application details and upcoming announcements. #ScopeTheScope 🔬 #MBL 🌊 #Microscopy 🔬 #BioImaging 🧬 #Fluorescence🪼#WoodsHole 🦞 #Optics 💡 #LightMicroscopy 🧪 #STEMCommunity 🫂 #Scientist

#AQLM Series 🧵: #Microscopy Mistakes -- Single Particle Tracking with Quantum Dots Single-particle tracking with quantum dots is highly sensitive to acquisition settings, labeling density, and localization precision. 🔬 📸 AQLM students learning SPT on a #Nikon AXR widefield system!

Analytical and Quantitative Light Microscopy Course

Analytical and Quantitative Light Microscopy Course

Mistake #3: Adding too many QDs. Over-labeling increases background and makes nearby particles difficult to separate. In dense regions, trajectories can overlap or swap identities during tracking.

Analytical and Quantitative Light Microscopy Course

Mistake #4: Assuming the Label Is Biologically Neutral #QuantumDots are physically large probes. Their size can alter receptor mobility or binding behavior, meaning the #fluorescent label itself may perturb the #biological process being measured. #MBL 🌊 #AQLM 🧫 #Microscopy 🔬 #BioImaging 🧬

Analytical and Quantitative Light Microscopy Course

Analytical and Quantitative Light Microscopy Course

Mistake #1: #Imaging too slowly. If frame rates are too low, rapidly moving particles appear to “jump” between positions, distorting trajectory reconstruction and diffusion analysis. #ScopeTheScope 🔬 #MBL 🌊 #AQLM 🧫 #Microscopy 🔬 #BioImaging 🧬 #Fluorescence🪼#WoodsHole 🦞 #Optics 💡 #LightMicroscopy 🧪

Analytical and Quantitative Light Microscopy Course

Mistake #2: #Imaging too quickly. Imaging too fast means each frame collects fewer #photons, reducing signal-to-noise and localization precision. Particles may appear dim, noisy, or intermittently lost during tracking. The optimal frame rate depends on the diffusion speed of your particle!

Analytical and Quantitative Light Microscopy Course

10 Sept

The Loeb Laboratory at the #MBL has been central to #research and training since its construction in 1970. MBL courses held in Loeb have trained hundreds of scientists each year and continue to shape modern biological research. 📸 Original photo #AQLM 🧫 #Microscopy 🔬 #Imaging🪼#WoodsHole 🦞 #Optics 💡

Analytical and Quantitative Light Microscopy Course

27 Aug

The Swope Center sits along Eel Pond, anchoring daily life at the #MBL. From meals and housing to informal discussions that extend beyond the lab, the Swope Center forms a space where science and community come together. #AQLM 🧫 #Microscopy 🔬 #WoodsHole 🦞 #STEMCommunity

Analytical and Quantitative Light Microscopy Course

13 Aug

The Lillie Building at the #MBL was established in 1925 as a purpose-designed space for delicate scientific work. A century later, it remains a place where #microscopy, collaboration, and new ideas come together. 📸 Original photo #AQLM 🧫 #BioImaging 🧬 #Fluorescence🪼#WoodsHole 🦞 #Optics 💡

#AQLM Series🧵: Microscopy Mistakes -- oh, FRAP! What happens after a #fluorescent molecule is bleached with a laser? In FRAP #microscopy, the recovery itself is the measurement. 🔬 📸 @3i Intelligent Imaging Innovations on the Marianas platform; 63× objective and a Photometrics Prime 95B camera. (1/7)

Analytical and Quantitative Light Microscopy Course

Analytical and Quantitative Light Microscopy Course

Mistake #4: Forgetting that recovery ≠ diffusion alone. A slow recovery curve may reflect binding interactions, compartmentalization, crowding, or active transport. --> Interpretation requires #biological context!! (6/7)

Analytical and Quantitative Light Microscopy Course

Mistake #5: Skipping normalization and proper controls. FRAP is #quantitative #microscopy. Recovery curves depend on #laser power, #imaging settings, ROI selection, drift correction, and reference regions. Small inconsistencies can completely change conclusions. (7/7)

Analytical and Quantitative Light Microscopy Course

Analytical and Quantitative Light Microscopy Course

Mistake #2: Ignoring acquisition photobleaching. Repeated #imaging slowly reduces #fluorescence across the sample, even outside the bleach spot. Without correction, recovery curves can underestimate #molecular mobility. (4/7)

Analytical and Quantitative Light Microscopy Course

Mistake #3: Bleaching too aggressively. Overbleaching can damage the sample, alter #molecular behavior, or increase background artifacts. The goal is measurable signal loss, so adjust your settings for your sample and #sceintific question. (5/7)

Analytical and Quantitative Light Microscopy Course

Analytical and Quantitative Light Microscopy Course

By tracking #fluorescence returning into the bleached region, FRAP can reveal #molecular mobility and binding dynamics in living #cells. But small experimental choices can dramatically alter recovery curves and change how the #data are interpreted. (2/7) #MBL 🌊 #AQLM 🧫 #Microscopy 🔬 #BioImaging 🧬

Analytical and Quantitative Light Microscopy Course

Mistake #1: Bleaching too large of a region. Large ROIs recover more slowly simply because #molecules must travel farther; recovery time is influenced by #geometry as well as #biology. (3/7)

Analytical and Quantitative Light Microscopy Course

13 July

Happy #MicroscopyMonday - #AQLM students doing single particle tracking using QD655 with THE Louise on a Leica DMi8 Inverted Widefield Microscope! Thank you for all your years at #AQLM, Louise! #ScopeTheScope 🔬 #MBL 🌊 #AQLM 🧫 #Microscopy 🔬 #BioImaging 🧬 #WoodsHole 🦞 #Optics 💡 #LightMicroscopy 🧪

Analytical and Quantitative Light Microscopy Course

10 July

Back by popular demand: Pets of AQLM, round two! Already looking ahead to the 2027 Pet Cohort. 🐾 #ScopeTheScope 🔬 #MBL 🌊 #AQLM 🧫 #Microscopy 🔬 #BioImaging 🧬 #Fluorescence🪼#WoodsHole 🦞 #Optics 💡 #LightMicroscopy 🧪 #FluorescenceMicroscopy ⭐️ #Confocal 🥼 #SuperResolution 🔥 #STEMCommunity 🫂

Analytical and Quantitative Light Microscopy Course

8 June

From Eel Pond, a community of #microscopic life comes into view. Captured in darkfield at 4X, 0.16 NA on an #Evident DP75 microscope, each branching form is defined by the light it scatters, revealing delicate edges and structure against a dark field. #ScopeTheScope 🔬 #MBL 🌊 #AQLM 🧫 #Microscopy 🔬

Analytical and Quantitative Light Microscopy Course

5 June

#ThrowbackThursday with #AQLM. Since its founding in 1980 by Shinya Inoué, AQLM has been closely tied to the evolution of #imaging living systems. Today, it continues to train new generations in quantitative biological imaging. #MBL 🌊 #Microscopy 🔬 #BioImaging 🧬 #WoodsHole 🦞 #Optics 💡#Confocal

Analytical and Quantitative Light Microscopy Course

10 May

Happy #MothersDay from this shrimp and her eggs, captured on an #Evident DP75 4X/0.16 NA darkfield microscope. #ScopeTheScope 🔬 #MBL 🌊 #AQLM 🧫 #Microscopy 🔬 #BioImaging 🧬 #Fluorescence🪼#WoodsHole 🦞 #Optics 💡 #LightMicroscopy

Analytical and Quantitative Light Microscopy Course

8 May

That’s a wrap on #AQLM 2026 🔬Thank you to everyone who made this year’s course possible: students, faculty, TAs and vendors. We are looking forward to what this #microscopy community builds next. Interested in applying next year? 2027 applications open in late 2026, stay tuned for updates.

Analytical and Quantitative Light Microscopy Course

8 May

It was an honor to hear Prof. Sarah Veatch, PhD, deliver yesterday’s Inoué Lecture, discussing how spatiotemporal pair-correlation analysis can recover nanoscale membrane organization in live-cell single-molecule localization #microscopy despite diffusion and molecular motion. #MBL 🌊 #AQLM 🧫

Analytical and Quantitative Light Microscopy Course reposted

Reto Fiolka

@retof.bsky.social

7 May

Back from @aqlm.bsky.social . Adequate sunset view on the way back.

Analytical and Quantitative Light Microscopy Course

7 May

Why be out of focus when you could be at #AQLM, learning about microscopes inSTED? Today, our students did rotations with our vendors #OxfordInstruments, #Nikon, #3i, #Leica & #ONI, learning about super-resolution #imaging modalities including DNA-PAINT and dSTORM. #MBL 🌊 #Microscopy 🔬

Analytical and Quantitative Light Microscopy Course

6 May

Good to have you back again this year, Jim! @thejimsims.bsky.social #MBL 🌊 #AQLM 🧫 #Microscopy 🔬 #WoodsHole 🦞 #Optics 💡 #LightMicroscopy 🔥 #STEMCommunity 🫂 #ScientistLife 🧑‍🔬

thejimsims.bsky.social@thejimsims.bsky.social

Analytical and Quantitative Light Microscopy Course

6 May

We tuned into a lecture by #AQLM course director, Prof. Diane Lidke, PhD, about single particle tracking. She discussed labelling strategies including quantum dots, and their applications using 2D/3D trajectory visualizations and diffusion mapping. #MBL 🌊 #Microscopy 🔬 #BioImaging 🧬 #Fluorescence🪼

6 May

Yesterday, we tuned into a lecture by Prof. Reto Fiolka, PhD, about Light-Sheet #Fluorescence #Microscopy. He discussed light sheet #optics and different #imaging modalities, as well as implementations in live imaging whole #embryos. #MBL 🌊 #AQLM 🧫 #BioImaging 🧬#WoodsHole 🦞 #LightMicroscopy 🧪

Analytical and Quantitative Light Microscopy Course reposted

Nat Prunet

@nat-prunet.bsky.social

Louse imaged on @zeiss-microscopy.bsky.social #lightsheet 7 #microscope by @carsten-wolff.bsky.social @mblscience.bsky.social @aqlm.bsky.social #microscopy

Analytical and Quantitative Light Microscopy Course reposted

Nat Prunet

@nat-prunet.bsky.social

@carsten-wolff.bsky.social teaching @aqlm.bsky.social students about light sheet #microscopy on the @zeiss-microscopy.bsky.social LightSheet 7. Feat. cute crustacean embryo for specimen

Analytical and Quantitative Light Microscopy Course reposted

Reto Fiolka

@retof.bsky.social

Analytical and Quantitative Light Microscopy Course

@rtomer.bsky.social : Pizza time at AQLM! @aqlm.bsky.social !

This sleek beam was created through a fluorescence cube on a Cairn OpenFrame, using a Toptica laser. ✨ #ScopeTheScope 🔬 #MBL 🌊 #AQLM 🧫 #Microscopy 🔬 #BioImaging 🧬 #Fluorescence🪼#WoodsHole 🦞 #Optics 💡 #LightMicroscopy 🧪 #FluorescenceMicroscopy ⭐️ #Confocal 🥼 #SuperResolution 🔥 #STEMCommunity 🫂

Analytical and Quantitative Light Microscopy Course reposted

Nat Prunet

@nat-prunet.bsky.social

It’s #lightsheet #microscopy day @aqlm.bsky.social with @retof.bsky.social

Analytical and Quantitative Light Microscopy Course reposted

Reto Fiolka

@retof.bsky.social

Analytical and Quantitative Light Microscopy Course

@aqlm.bsky.social Quite reasonable requests…

Analytical and Quantitative Light Microscopy Course

Nickelback said to "look at this graph": A rail-mounted light sheet #microscope creating a sharp intensity peak. So cool! #ScopeTheScope 🔬 #MBL 🌊 #AQLM 🧫 #Microscopy 🔬 #BioImaging 🧬 #Fluorescence🪼#WoodsHole 🦞 #Optics 💡 #LightMicroscopy 🧪 #FluorescenceMicroscopy ⭐️ #Confocal 🥼 #SuperResolution 🔥

Analytical and Quantitative Light Microscopy Course reposted

@the.3i.social

4 May

We are @mblscience.bsky.social @aqlm.bsky.social sharing the latest live cell imaging technology with our Marianas SDC system. 3i's LaserStack, Vector3 TIRF/FRAP, and components from ...

Analytical and Quantitative Light Microscopy Course

4 May

Real-time imaging at #AQLM, where “woooo got a signal” is a totally valid reaction! 🔬 #ScopeTheScope 🔬 #MBL 🌊 #AQLM 🧫 #Microscopy 🔬 #BioImaging 🧬 #Fluorescence🪼#WoodsHole 🦞 #Optics 💡 #LightMicroscopy 🧪 #FluorescenceMicroscopy ⭐️ #Confocal 🥼 #SuperResolution 🔥 #STEMCommunity 🫂 #ScientistLife 🧑‍🔬