scholarly journals Facile assembly of an affordable miniature multicolor fluorescence microscope made of 3D-printed parts enables detection of single cells

2019 ◽  
Author(s):  
Samuel B. Tristan-Landin ◽  
Alan M. Gonzalez-Suarez ◽  
Rocio J. Jimenez-Valdes ◽  
Jose L. Garcia-Cordero
Keyword(s):  
Biological Process ◽  
Single Cells ◽  
Time Lapse ◽  
Laboratory Diagnosis ◽  
Fluorescence Microscope ◽  
Electronic Components ◽  
Bright Field ◽  
Field Mode ◽  
3D Printed ◽  
Multicolor Fluorescence

AbstractFluorescence microscopy is one of the workhorses of biomedical research and laboratory diagnosis; however, their cost, size, maintenance, and fragility has prevented their adoption in developing countries or low-resource settings. Although significant advances have decreased their size, cost and accessibility, their designs and assembly remain rather complex. Here, inspired on the simple mechanism from a nut and a bolt we report the construction of a portable fluorescence microscope that operates in bright field mode and in three fluorescence channels: UV, green, and red. It is assembled in under 10 min from only six 3D printed parts and basic electronic components that can be readily purchased in most locations or online for US $85. Adapting a microcomputer and a touch LCD screen, the microscope can capture time-lapse images and videos. We characterized its resolution and illumination conditions and benchmarked its performance against a high-end fluorescence microscope by tracking a biological process in single cells. We also demonstrate its application to image cells inside a microfluidic device in bright-field and fluorescence mode. Our microscope fits in a CO2 chamber and can be operated in time-lapse mode. Our portable microscope is ideal in applications where space is at a premium, such as lab-on-a-chips or space missions, and can find applications in clinical research, diagnostics, telemedicine and in educational settings.

scholarly journals Facile assembly of an affordable miniature multicolor fluorescence microscope made of 3D-printed parts enables detection of single cells

PLoS ONE ◽  
2019 ◽  
Vol 14 (10) ◽  
pp. e0215114 ◽  
Author(s):  
Samuel B. Tristan-Landin ◽  
Alan M. Gonzalez-Suarez ◽  
Rocio J. Jimenez-Valdes ◽  
Jose L. Garcia-Cordero
Keyword(s):  
Single Cells ◽  
Fluorescence Microscope ◽  
3D Printed ◽  
Multicolor Fluorescence

Imaging of platinum-shadowed macromolecular complexes in dark field

1984 ◽  
Vol 42 ◽  
pp. 146-147
Author(s):  
D.W. Andrews ◽  
F.P. Ottensmeyer
Keyword(s):  
Thin Film ◽  
Three Dimensional ◽  
Average Diameter ◽  
Dark Field ◽  
Bright Field ◽  
Field Mode ◽  
Macromolecular Complexes ◽  
Average Mass ◽  
Topological Features ◽  
Projection Images

Shadowing with heavy metals has been used for many years to enhance the topological features of biological macromolecular complexes. The three dimensional features present in directionaly shadowed specimens often simplifies interpretation of projection images provided by other techniques. One difficulty with the method is the relatively large amount of metal used to achieve sufficient contrast in bright field images. Thick shadow films are undesirable because they decrease resolution due to an increased tendency for microcrystalline aggregates to form, because decoration artefacts become more severe and increased cap thickness makes estimation of dimensions more uncertain.The large increase in contrast provided by the dark field mode of imaging allows the use of shadow replicas with a much lower average mass thickness. To form the images in Fig. 1, latex spheres of 0.087 μ average diameter were unidirectionally shadowed with platinum carbon (Pt-C) and a thin film of carbon was indirectly evaporated on the specimen as a support.

On the cross-sectional shape of the cellulose crystallites in the tunicate Halocynthia papillosa

1990 ◽  
Vol 48 (3) ◽  
pp. 566-567
Author(s):  
J.-F. Revol ◽  
Y. Van Daele ◽  
F. Gaill
Keyword(s):  
Contrast Imaging ◽  
Animal Kingdom ◽  
Bright Field ◽  
Field Mode ◽  
Cross Sectional ◽  
Ultrathin Sections ◽  
The Cross ◽  
Sectional Shape ◽  
Valonia Ventricosa ◽  
C Nmr

The only form of cellulose which could unequivocally be ascribed to the animal kingdom is the tunicin that occurs in the tests of the tunicates. Recently, high-resolution solid-state l3C NMR revealed that tunicin belongs to the Iβ form of cellulose as opposed to the Iα form found in Valonia and bacterial celluloses. The high perfection of the tunicin crystallites led us to study its crosssectional shape and to compare it with the shape of those in Valonia ventricosa (V.v.), the goal being to relate the cross-section of cellulose crystallites with the two allomorphs Iα and Iβ.In the present work the source of tunicin was the test of the ascidian Halocvnthia papillosa (H.p.). Diffraction contrast imaging in the bright field mode was applied on ultrathin sections of the V.v. cell wall and H.p. test with cellulose crystallites perpendicular to the plane of the sections. The electron microscope, a Philips 400T, was operated at 120 kV in a low intensity beam condition.

scholarly journals 3D printed microfluidic lab-on-a-chip device for fiber-based dual beam optical manipulation

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Haoran Wang ◽  
Anton Enders ◽  
John-Alexander Preuss ◽  
Janina Bahnemann ◽  
Alexander Heisterkamp ◽  
...  
Keyword(s):  
Optical Fibers ◽  
Single Cells ◽  
Lab On A Chip ◽  
Cost Effective ◽  
Optical Manipulation ◽  
Hydrodynamic Focusing ◽  
Trapping Region ◽  
Dual Beam ◽  
3D Printed ◽  
On Chip

Abstract3D printing of microfluidic lab-on-a-chip devices enables rapid prototyping of robust and complex structures. In this work, we designed and fabricated a 3D printed lab-on-a-chip device for fiber-based dual beam optical manipulation. The final 3D printed chip offers three key features, such as (1) an optimized fiber channel design for precise alignment of optical fibers, (2) an optically clear window to visualize the trapping region, and (3) a sample channel which facilitates hydrodynamic focusing of samples. A square zig–zag structure incorporated in the sample channel increases the number of particles at the trapping site and focuses the cells and particles during experiments when operating the chip at low Reynolds number. To evaluate the performance of the device for optical manipulation, we implemented on-chip, fiber-based optical trapping of different-sized microscopic particles and performed trap stiffness measurements. In addition, optical stretching of MCF-7 cells was successfully accomplished for the purpose of studying the effects of a cytochalasin metabolite, pyrichalasin H, on cell elasticity. We observed distinct changes in the deformability of single cells treated with pyrichalasin H compared to untreated cells. These results demonstrate that 3D printed microfluidic lab-on-a-chip devices offer a cost-effective and customizable platform for applications in optical manipulation.

scholarly journals Stochasticity in Colonial Growth Dynamics of Individual Bacterial Cells

2013 ◽  
Vol 79 (7) ◽  
pp. 2294-2301 ◽  
Author(s):  
Konstantinos P. Koutsoumanis ◽  
Alexandra Lianou
Keyword(s):  
Kinetic Parameters ◽  
Single Cells ◽  
Growth Curves ◽  
Growth Dynamics ◽  
Time Lapse ◽  
Initial Population ◽  
Bacterial Cells ◽  
Stochastic Growth ◽  
Bacterial Populations ◽  
Content Type

ABSTRACTConventional bacterial growth studies rely on large bacterial populations without considering the individual cells. Individual cells, however, can exhibit marked behavioral heterogeneity. Here, we present experimental observations on the colonial growth of 220 individual cells ofSalmonella entericaserotype Typhimurium using time-lapse microscopy videos. We found a highly heterogeneous behavior. Some cells did not grow, showing filamentation or lysis before division. Cells that were able to grow and form microcolonies showed highly diverse growth dynamics. The quality of the videos allowed for counting the cells over time and estimating the kinetic parameters lag time (λ) and maximum specific growth rate (μmax) for each microcolony originating from a single cell. To interpret the observations, the variability of the kinetic parameters was characterized using appropriate probability distributions and introduced to a stochastic model that allows for taking into account heterogeneity using Monte Carlo simulation. The model provides stochastic growth curves demonstrating that growth of single cells or small microbial populations is a pool of events each one of which has its own probability to occur. Simulations of the model illustrated how the apparent variability in population growth gradually decreases with increasing initial population size (N0). For bacterial populations withN0of >100 cells, the variability is almost eliminated and the system seems to behave deterministically, even though the underlying law is stochastic. We also used the model to demonstrate the effect of the presence and extent of a nongrowing population fraction on the stochastic growth of bacterial populations.

scholarly journals Measuring fast gene dynamics in single cells with time-lapse luminescence microscopy

2014 ◽  
Vol 25 (22) ◽  
pp. 3699-3708 ◽  
Author(s):  
Anyimilehidi Mazo-Vargas ◽  
Heungwon Park ◽  
Mert Aydin ◽  
Nicolas E. Buchler
Keyword(s):  
Gene Expression ◽  
Fluorescent Proteins ◽  
Single Cells ◽  
Time Lapse ◽  
Photon Flux ◽  
Luminescence Microscopy ◽  
Cell Cycle Genes ◽  
Light Excitation ◽  
Better Than

Time-lapse fluorescence microscopy is an important tool for measuring in vivo gene dynamics in single cells. However, fluorescent proteins are limited by slow chromophore maturation times and the cellular autofluorescence or phototoxicity that arises from light excitation. An alternative is luciferase, an enzyme that emits photons and is active upon folding. The photon flux per luciferase is significantly lower than that for fluorescent proteins. Thus time-lapse luminescence microscopy has been successfully used to track gene dynamics only in larger organisms and for slower processes, for which more total photons can be collected in one exposure. Here we tested green, yellow, and red beetle luciferases and optimized substrate conditions for in vivo luminescence. By combining time-lapse luminescence microscopy with a microfluidic device, we tracked the dynamics of cell cycle genes in single yeast with subminute exposure times over many generations. Our method was faster and in cells with much smaller volumes than previous work. Fluorescence of an optimized reporter (Venus) lagged luminescence by 15–20 min, which is consistent with its known rate of chromophore maturation in yeast. Our work demonstrates that luciferases are better than fluorescent proteins at faithfully tracking the underlying gene expression.

scholarly journals Soldering of Electronics Components on 3D-Printed Conductive Substrates

Materials ◽  
2021 ◽  
Vol 14 (14) ◽  
pp. 3850
Author(s):  
Bartłomiej Podsiadły ◽  
Andrzej Skalski ◽  
Marcin Słoma
Keyword(s):  
Additive Manufacturing ◽  
Solder Joints ◽  
Rapid Development ◽  
Surface Preparation ◽  
Contact Angles ◽  
Electronic Components ◽  
Optimal Method ◽  
Structural Electronics ◽  
3D Printed ◽  
Composite Substrates

Rapid development of additive manufacturing and new composites materials with unique properties are promising tools for fabricating structural electronics. However, according to the typical maximum resolution of additive manufacturing methods, there is no possibility to fabricate all electrical components with these techniques. One way to produce complex structural electronic circuits is to merge 3D-printed elements with standard electronic components. Here, different soldering and surface preparation methods before soldering are tested to find the optimal method for soldering typical electronic components on conductive, 3D-printed, composite substrates. To determine the optimal soldering condition, the contact angles of solder joints fabricated in different conditions were measured. Additionally, the mechanical strength of the joints was measured using the shear force test. The research shows a possibility of fabricating strong, conductive solder joints on composites substrates prepared by additive manufacturing. The results show that mechanical cleaning and using additional flux on the composite substrates are necessary to obtain high-quality solder joints. The most repeatable joints with the highest shear strength values were obtained using reflow soldering together with low-temperature SnBiAg solder alloy. A fabricated demonstrator is a sample of the successful merging of 3D-printed structural electronics with standard electronic components.

Growth in cell length in the fission yeast Schizosaccharomyces pombe

1985 ◽  
Vol 75 (1) ◽  
pp. 357-376 ◽  
Author(s):  
J.M. Mitchison ◽  
P. Nurse
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Cell Size ◽  
Single Cells ◽  
Temperature Shift ◽  
Time Lapse ◽  
Cell Length ◽  
Wild Type ◽  
Cycle Growth ◽  
A Cell ◽  
Mutant Cells

The cylindrical cells of Schizosaccharomyces pombe grow in length by extension at the ends and not the middle. At the beginning of the cell cycle, growth is restricted to the ‘old end’, which existed in the previous cycle. Later on, the ‘new end’, formed from the septum, starts to grow at a point in the cycle that we have called NETO (‘new end take-off’). Fluorescence microscopy on cells stained with Calcofluor has been used to study NETO in size mutants, in blocked cdc mutants and with different growth temperatures and media. In wild-type cells (strain 972) NETO happens at 0.34 of the cycle with a cell length of 9.5 microns. With size mutants that are smaller at division, NETO takes place at the same size (9.0-9.5 microns) but this is not achieved until later in the cycle. Another control operates in larger size mutants since NETO occurs at the same stage of the cycle (about 0.32) as in wild type but at a larger cell size. This control is probably a requirement to have completed an event in early G2, since most cdc mutant cells blocked before this point in the cycle do not show NETO whereas most of those blocked in late G2 do show it. We conclude that NETO only happens if: (1) the cell length is greater than a critical value of 9.0-9.5 microns; and (2) the cell has traversed the first 0.3-0.35 of the cycle and passed early G2. NETO is delayed in poor media, in which cell size is also reduced. Temperature has little effect on NETO under steady-state conditions, but there is a transient delay for some hours after a temperature shift. NETO is later in another wild-type strain, 132. Time-lapse photomicrography was used to follow the rates of length growth in single cells. Wild-type cells showed two linear segments during the first 75% of the cycle. There was a rate-change point (RCP), coincident with NETO, where the rate of total length extension increased by 35%. This increase was not due simply to the start of new-end growth, since old-end growth slowed down in some cells at the RCP. cdc 11.123 is a mutant in which septation and division is blocked at 35 degrees C but nuclear division continues.(ABSTRACT TRUNCATED AT 400 WORDS)

Fluids—Good and Bad Actors: Observations From the College of American Pathologists Interlaboratory Comparison Program in Nongynecologic Cytology

2004 ◽  
Vol 128 (5) ◽  
pp. 513-518
Author(s):  
Ann T. Moriarty ◽  
Janet Stastny ◽  
Emily E. Volk ◽  
Jonathan H. Hughes ◽  
Theodore R. Miller ◽  
...  
Keyword(s):  
Cell Carcinoma ◽  
Interlaboratory Comparison ◽  
Single Cells ◽  
Poor Performance ◽  
Laboratory Diagnosis ◽  
Body Cavity ◽  
Lymphoid Cells ◽  
Performance Characteristics ◽  
Careful Examination ◽  
Diagnostic Challenge

Abstract Context.—Body cavity fluid examination presents a common and sometimes difficult diagnostic challenge in daily cytology practice. Separating benign from malignant cellular changes may require meticulous screening, careful scrutiny of cellular features, and an understanding of the range of reactive changes. We use the data from the College of American Pathologists (CAP) Interlaboratory Comparison Program in Nongynecologic Cytology (NGC) to identify characteristics of fluids that place them at opposite ends of the diagnostic spectrum. Objective.—To assess the features of individual body cavity fluid slides that demonstrated good performance characteristics and compare them to slides that were poor performers. Design.—A databank of 10 396 laboratory responses, including a variety of malignant and benign cases obtained from 1997 through 2001, was used to select cases. A cumulative slide history was used to identify slides that performed well or poorly in each reference diagnosis. Cases were confirmed by consensus of 4 CAP Cytopathology Resource Committee members. Observations and characterizations of good and bad performers in each category were recorded and summarized. Results.—Percentage of concordance of poor performers ranged from 0% to 58%. Conversely, good performers were identified with high concordance of laboratory diagnosis in each reference category (>80%). Several patterns emerged. Poorly performing cases of adenocarcinoma consisted of slides with rare tumor cells, hypercellular malignant cases without 2 cell populations, and cases with single cells. Poor performance in confirmed squamous cell carcinoma cases related to rare cells without keratinization. Small cell carcinoma and melanoma cases performed poorly when there were few malignant cells. Lymphoma cases demonstrated poor performance when there were abundant pleomorphic lymphoid cells or when rare Reed- Sternberg–like cells were present. Reactive or negative slides performed best with a polymorphous population; poor performers were those with a predominant lymphocyte population mistaken for a hematopoietic neoplasm. Conclusion.—Close attention to classic cytologic criteria and careful examination of slides may enhance the educational experience of participants and the performance characteristics of body cavity fluid specimens in the CAP NGC program. Lessons from bad actors in the CAP NGC program may increase awareness of potential diagnostic problems in daily practice or help identify areas for laboratory quality improvement.

Sign in / Sign up

Export Citation Format

Share Document