Droplet-based light-sheet fluorescence microscopy for high-throughput sample preparation, 3-D imaging and quantitative analysis on a chip

Lab on a Chip ◽  
2017 ◽  
Vol 17 (13) ◽  
pp. 2193-2197 ◽  
Author(s):  
Hao Jiang ◽  
Tingting Zhu ◽  
Hao Zhang ◽  
Jun Nie ◽  
Zeyi Guan ◽  
...  
Keyword(s):  
Quantitative Analysis ◽  
Fluorescence Microscopy ◽  
Sample Preparation ◽  
High Throughput ◽  
Light Sheet ◽  
Droplet Microfluidics ◽  
On Chip ◽  
Light Sheet Fluorescence Microscopy

An optofluidic device interfaces droplet microfluidics with light-sheet fluorescence microscopy for on-chip, high-throughput sample compartmentalization and 3-D imaging.

scholarly journals High-throughput 3D imaging of single cells with light-sheet fluorescence microscopy on chip

2020 ◽  
Vol 11 (8) ◽  
pp. 4397 ◽  
Author(s):  
Federico Sala ◽  
Michele Castriotta ◽  
Petra Paiè ◽  
Andrea Farina ◽  
Sarah D’Annunzio ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
High Throughput ◽  
3D Imaging ◽  
Single Cells ◽  
Light Sheet ◽  
On Chip ◽  
Light Sheet Fluorescence Microscopy

scholarly journals Automatic imaging of Drosophila embryos with light sheet fluorescence microscopy on chip

2020 ◽  
Author(s):  
Roberto Memeo ◽  
Petra Paiè ◽  
Federico Sala ◽  
Michele Castriotta ◽  
Chiara Guercio ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Light Sheet ◽  
On Chip ◽  
Light Sheet Fluorescence Microscopy ◽  
Drosophila Embryos

scholarly journals Automated High-Throughput Light-Sheet Fluorescence Microscopy of Larval Zebrafish

2018 ◽  
Author(s):  
Savannah L. Logan ◽  
Christopher Dudley ◽  
Ryan P. Baker ◽  
Michael J. Taormina ◽  
Edouard A. Hay ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
High Throughput ◽  
Three Dimensional ◽  
Light Sheet ◽  
Individual Variability ◽  
Three Dimensional Imaging ◽  
Larval Zebrafish ◽  
Neutrophil Number ◽  
Spatially Resolved ◽  
Light Sheet Fluorescence Microscopy

ABSTRACTLight sheet fluorescence microscopy enables fast, minimally phototoxic, three-dimensional imaging of live specimens, but is currently limited by low throughput and tedious sample preparation. Here, we describe an automated high-throughput light sheet fluorescence microscope in which specimens are positioned by and imaged within a fluidic system integrated with the sheet excitation and detection optics. We demonstrate the ability of the instrument to rapidly examine live specimens with minimal manual intervention by imaging fluorescent neutrophils over a nearly 0.3 mm3 volume in dozens of larval zebrafish. In addition to revealing considerable inter-individual variability in neutrophil number, known previously from labor-intensive methods, three-dimensional imaging allows assessment of the correlation between the bulk measure of total cellular fluorescence and the spatially resolved measure of actual neutrophil number per animal. We suggest that our simple experimental design should considerably expand the scope and impact of light sheet imaging in the life sciences.

scholarly journals Automated high-throughput light-sheet fluorescence microscopy of larval zebrafish

PLoS ONE ◽  
2018 ◽  
Vol 13 (11) ◽  
pp. e0198705 ◽  
Author(s):  
Savannah L. Logan ◽  
Christopher Dudley ◽  
Ryan P. Baker ◽  
Michael J. Taormina ◽  
Edouard A. Hay ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
High Throughput ◽  
Light Sheet ◽  
Larval Zebrafish ◽  
Light Sheet Fluorescence Microscopy

scholarly journals Virtual unfolding of light sheet fluorescence microscopy dataset for quantitative analysis of the mouse intestine

2016 ◽  
Vol 21 (05) ◽  
pp. 1 ◽  
Author(s):  
Alessia Candeo ◽  
Luigi Maiuri ◽  
Ilenia Sana ◽  
Eleonora Ferrari ◽  
Cosimo D’Andrea ◽  
...  
Keyword(s):  
Quantitative Analysis ◽  
Fluorescence Microscopy ◽  
Light Sheet ◽  
Mouse Intestine ◽  
Light Sheet Fluorescence Microscopy

scholarly journals Cytotoxic effects and tolerability of gemcitabine and axitinib in a xenograft model for c-myc amplified medulloblastoma

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Stefanie Schwinn ◽  
Zeinab Mokhtari ◽  
Sina Thusek ◽  
Theresa Schneider ◽  
Anna-Leena Sirén ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Tumor Growth ◽  
Intensive Therapy ◽  
Xenograft Model ◽  
Light Sheet ◽  
Tumor Control ◽  
Orthotopic Model ◽  
Group 3 ◽  
Light Sheet Fluorescence Microscopy

AbstractMedulloblastoma is the most common high-grade brain tumor in childhood. Medulloblastomas with c-myc amplification, classified as group 3, are the most aggressive among the four disease subtypes resulting in a 5-year overall survival of just above 50%. Despite current intensive therapy regimens, patients suffering from group 3 medulloblastoma urgently require new therapeutic options. Using a recently established c-myc amplified human medulloblastoma cell line, we performed an in-vitro-drug screen with single and combinatorial drugs that are either already clinically approved or agents in the advanced stage of clinical development. Candidate drugs were identified in vitro and then evaluated in vivo. Tumor growth was closely monitored by BLI. Vessel development was assessed by 3D light-sheet-fluorescence-microscopy. We identified the combination of gemcitabine and axitinib to be highly cytotoxic, requiring only low picomolar concentrations when used in combination. In the orthotopic model, gemcitabine and axitinib showed efficacy in terms of tumor control and survival. In both models, gemcitabine and axitinib were better tolerated than the standard regimen comprising of cisplatin and etoposide phosphate. 3D light-sheet-fluorescence-microscopy of intact tumors revealed thinning and rarefication of tumor vessels, providing one explanation for reduced tumor growth. Thus, the combination of the two drugs gemcitabine and axitinib has favorable effects on preventing tumor progression in an orthotopic group 3 medulloblastoma xenograft model while exhibiting a favorable toxicity profile. The combination merits further exploration as a new approach to treat high-risk group 3 medulloblastoma.

Investigating safe excitation in Light Sheet Fluorescence Microscopy

2021 ◽  
Vol 84 ◽  
pp. 296
Author(s):  
Gideon Oluniran ◽  
James Blackwell ◽  
Emmanuel Reynaud ◽  
Marcin Krasny ◽  
Niall Colgan ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Light Sheet ◽  
Light Sheet Fluorescence Microscopy

scholarly journals Light sheet fluorescence microscopy of optically cleared brains for studying the glymphatic system

2020 ◽  
Vol 40 (10) ◽  
pp. 1975-1986
Author(s):  
Nicholas B Bèchet ◽  
Tekla M Kylkilahti ◽  
Bengt Mattsson ◽  
Martina Petrasova ◽  
Nagesh C Shanbhag ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Fluid Transport ◽  
Light Sheet ◽  
Brain Parenchyma ◽  
Sleep State ◽  
Glymphatic System ◽  
Light Sheet Microscopy ◽  
Highly Active ◽  
Quantitative Analyses ◽  
Light Sheet Fluorescence Microscopy

Fluid transport in the perivascular space by the glia-lymphatic (glymphatic) system is important for the removal of solutes from the brain parenchyma, including peptides such as amyloid-beta which are implicated in the pathogenesis of Alzheimer’s disease. The glymphatic system is highly active in the sleep state and under the influence of certain of anaesthetics, while it is suppressed in the awake state and by other anaesthetics. Here we investigated whether light sheet fluorescence microscopy of whole optically cleared murine brains was capable of detecting glymphatic differences in sleep- and awake-mimicking anaesthesia, respectively. Using light-sheet imaging of whole brains, we found anaesthetic-dependent cerebrospinal fluid (CSF) influx differences, including reduced tracer influx along tertiary branches of the middle cerebral artery and reduced influx along dorsal and anterior penetrating arterioles, in the awake-mimicking anaesthesia. This study establishes that light sheet microscopy of optically cleared brains is feasible for quantitative analyses and can provide images of the entire glymphatic system in whole brains.

FEATURES

2016 ◽  
Vol 20 (12) ◽  
pp. 12-22
Keyword(s):  
Medical Imaging ◽  
Fluorescence Microscopy ◽  
Fluorescence Spectroscopy ◽  
Medical Images ◽  
Light Sheet ◽  
The Future ◽  
Light Sheet Fluorescence Microscopy

Scanning the Future of Medical Imaging Putting Numbers into Biology: The Combination of Light Sheet Fluorescence Microscopy and Fluorescence Spectroscopy Abyss Processing – Exploring the Deep in Medical Images

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