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Microscope in Action: An Interdisciplinary Fluorescence Microscopy Hands-on Resource for Schools
G. Paci,
E. Haas,
L. Kornau,
D. Marchetti,
L. Wang,
R. Prevedel, and
A. Szmolenszky
Article Category: Research Article
Volume/Issue: Volume 2: Issue 3
Online Publication Date: Oct 07, 2021
DOI: 10.35459/tbp.2020.000171
Page Range: 55 – 73

I. INTRODUCTION Fluorescence is a phenomenon ubiquitous in nature that occurs when molecules (fluorophores) absorb light of a specific wavelength and emit light at a different (typically longer) wavelength. By staining cellular structures of interest with fluorescent dyes of distinct colors, fluorescence microscopy enables the visualization of molecules with a high degree of specificity. This powerful tool has completely revolutionized biology, and it represents a perfect example of the advancements enabled by biophysical research, as

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Designing a Compact, Low-Cost FRET Microscopy Platform for the Undergraduate Classroom
John W. Rupel,
Sophia M. Sdao,
Kadina E. Johnston,
Ethan T. Nethery,
Kaitlyn A. Gabardi,
Benjamin A. Ratliff,
Zach J. Simmons,
Jack T. Postlewaite,
Angela M. Kita,
Jeremy D. Rogers, and
Matthew J. Merrins
Article Category: Research Article
Volume/Issue: Volume 1: Issue 2
Online Publication Date: Jun 03, 2020
DOI: 10.35459/tbp.2019.000117
Page Range:

I. INTRODUCTION Fluorescence microscopy is a powerful technique that has revolutionized the field of biology by providing a relatively noninvasive approach to study cellular dynamics. A large selection of fluorescent biosensors is currently available that includes epitope tags for synthetic fluorophores ( 1 – 3 ), dyes ( 4 – 6 ), and genetically encoded sensors ( 7 – 9 ). Förster resonance energy transfer (FRET)–based sensors ( 10 – 13 ) are still the most prevalent and diverse, having been leveraged to measure protein–protein interactions

Fig 5; Fraction of correctly answered questions on subjects of natural sciences and microscopy, before and after using the LEGO microscope. The bars show the average results of 5 questions on either of the subjects, filled in by 8 students in the age range between 9 and 13 y. The black dots correspond to the results of individual students. There was no significant (ns) change in the results of the general science questions: from 78 ± 21% to 83 ± 21% correctly answered questions, but a statistically significant improvement (***, P < 0.001) was observed for the questions on microscopy, from 50 ± 17% to 83 ± 12% correct answers.
Bart E. Vos,
Emil Betz Blesa, and
Timo Betz
Fig 5
Fig 5

Fraction of correctly answered questions on subjects of natural sciences and microscopy, before and after using the LEGO microscope. The bars show the average results of 5 questions on either of the subjects, filled in by 8 students in the age range between 9 and 13 y. The black dots correspond to the results of individual students. There was no significant (ns) change in the results of the general science questions: from 78 ± 21% to 83 ± 21% correctly answered questions, but a statistically significant improvement (***, P < 0.001) was observed for the questions on microscopy, from 50 ± 17% to 83 ± 12% correct answers.

Designing a High-Resolution, LEGO-Based Microscope for an Educational Setting
Bart E. Vos,
Emil Betz Blesa, and
Timo Betz
Article Category: Research Article
Volume/Issue: Volume 2: Issue 3
Online Publication Date: Jun 22, 2021
DOI: 10.35459/tbp.2021.000191
Page Range: 29 – 40

I. INTRODUCTION The invention of microscopy in the 17th century by Antoni van Leeuwenhoek was the start of an era of research in the micro-world ( 1 ). Although we know by now what the “little animals” are that he observed, the micro- and nano-world is an inexhaustible topic for biophysical research, for which the light microscope is the instrument of choice. Despite the simplicity of a basic light microscope, the fundamental working principles are beyond everyday intuition for pupils, but also for most adults. Although this lack of

Hands-on Curriculum in Optics of Microscopy
Rachel Kemp,
Alexander Chippendale,
Monica Harrelson,
Jennifer Shumway,
Amanda Tan,
Sarah Zuraw, and
Jennifer L. Ross
Article Category: Research Article
Volume/Issue: Volume 1: Issue 1
Online Publication Date: Feb 14, 2020
DOI: 10.35459/tbp.2019.000114
Page Range:

I. INTRODUCTION Optics is the study of how light interacts with matter. For centuries, people have been able to manipulate materials to bend and reflect light. For the past 800 years, humans have shaped lenses from high-index materials to enable magnified imaging of objects. The combination of lenses into instruments resembling microscopes dates back 500 years. Despite the long history of microscopic measurement, advances in microscopy techniques are still being made today. For instance, the 2014 Nobel Prize in Chemistry was awarded for

A Microscope Medley for High School Students
Raghuveer Parthasarathy
Article Category: Research Article
Volume/Issue: Volume 5: Issue 1
Online Publication Date: Jul 04, 2024
DOI: 10.35459/tbp.2024.000261
Page Range: 47 – 54

I. INTRODUCTION The awe-inspiring ability of microscopes to make visible the world of very small things is well known. In contrast, the variety of approaches to microscopy, and even the variety of aims of microscopy, are not well known either by the public in general or by secondary school (high school) students in particular. In addition to constraining the appreciation of contemporary science and technology, this lack of awareness restricts students’ views of careers and paths of study. The relevance of microscopy to biophysics has been evident since

Fig 10; Biophysical application: Notochord of the zebrafish under Koehler and critical illuminations. The images were taken by light sheet fluorescence microscopy (LSFM) with a 40× 0.8 numerical aperture (NA) water immersion objective. (A) The 4f system with full NA. (B) An LED flashlight in direct illumination. (C, D) Regions within the dashed rectangle in panels A and B, respectively. Skeletal muscle fibers are distinguishable in panel C, but hardly in panel D.
Jorge Madrid-Wolff and
Manu Forero-Shelton
Fig 10
Fig 10

Biophysical application: Notochord of the zebrafish under Koehler and critical illuminations. The images were taken by light sheet fluorescence microscopy (LSFM) with a 40× 0.8 numerical aperture (NA) water immersion objective. (A) The 4f system with full NA. (B) An LED flashlight in direct illumination. (C, D) Regions within the dashed rectangle in panels A and B, respectively. Skeletal muscle fibers are distinguishable in panel C, but hardly in panel D.

Fig 2; A home-built light sheet fluorescence microscope. (a) Lenses, mirrors, and other optical elements. Laser illumination (blue) is shaped into a thin sheet and directed to the specimen chamber, near the bottom of the image. (b) The specimen chamber. Larval zebrafish are held in plugs of agar gel suspended from the ends of glass capillaries. (c) Illustration of the laser sheet orientation (blue) based on the image in (b). The sheet is not drawn to scale; its vertical extent is roughly 0.4 mm to match the detection field of view.
Raghuveer Parthasarathy
Fig 2
Fig 2

A home-built light sheet fluorescence microscope. (a) Lenses, mirrors, and other optical elements. Laser illumination (blue) is shaped into a thin sheet and directed to the specimen chamber, near the bottom of the image. (b) The specimen chamber. Larval zebrafish are held in plugs of agar gel suspended from the ends of glass capillaries. (c) Illustration of the laser sheet orientation (blue) based on the image in (b). The sheet is not drawn to scale; its vertical extent is roughly 0.4 mm to match the detection field of view.

Fig 3; Example light sheet microscope images of a larval zebrafish with fluorescent immune cells. (a) Schematic illustration of a 6-d postfertilization zebrafish larva. The swim bladder is outlined in cyan, and the gut is outlined in magenta. The orange box indicates the region captured by the field of view in the subsequent panels. Scale bar: 0.5 mm. (b) Bright-field image of part of a larval zebrafish. Scale bar: 0.1 mm. (c and d) The 2 light sheet fluorescence microscope images, from planes separated by 23 μm and from the same fish and the same field of view as in (b), showing fluorescent neutrophils in this transgenic fish. Insets are expanded by 3×. The neutrophil in the plane shown in (d), located in a fin, can be discerned in the bright-field image, but the neutrophils in (c), situated in thick, dense tissue near the gut, cannot.
Raghuveer Parthasarathy
Fig 3
Fig 3

Example light sheet microscope images of a larval zebrafish with fluorescent immune cells. (a) Schematic illustration of a 6-d postfertilization zebrafish larva. The swim bladder is outlined in cyan, and the gut is outlined in magenta. The orange box indicates the region captured by the field of view in the subsequent panels. Scale bar: 0.5 mm. (b) Bright-field image of part of a larval zebrafish. Scale bar: 0.1 mm. (c and d) The 2 light sheet fluorescence microscope images, from planes separated by 23 μm and from the same fish and the same field of view as in (b), showing fluorescent neutrophils in this transgenic fish. Insets are expanded by 3×. The neutrophil in the plane shown in (d), located in a fin, can be discerned in the bright-field image, but the neutrophils in (c), situated in thick, dense tissue near the gut, cannot.

Fig 1; (a) One of Antonie van Leeuwenhoek’s early microscopes (photo by Jacopo Werther, licensed under the Creative Commons Attribution-ShareAlike 3.0 Unported license; https://en.m.wikipedia.org/wiki/File:Leeuwenhoek_Microscope.png). (b) A simple transmitted-light compound microscope. (c) Color-printed letters with a ruler for scale, as viewed through the microscope. The yellow and cyan inks making up the green “o” are evident.
Raghuveer Parthasarathy
Fig 1
Fig 1

(a) One of Antonie van Leeuwenhoek’s early microscopes (photo by Jacopo Werther, licensed under the Creative Commons Attribution-ShareAlike 3.0 Unported license; https://en.m.wikipedia.org/wiki/File:Leeuwenhoek_Microscope.png). (b) A simple transmitted-light compound microscope. (c) Color-printed letters with a ruler for scale, as viewed through the microscope. The yellow and cyan inks making up the green “o” are evident.