Vascular organ-on-chip imaging: a dedicated single plane illumination fluorescence microscope

Miniaturizing flow cytometry requires a comprehensive approach to redesigning the conventional fluidic and optical systems to have a small footprint and simple usage and to enable rapid cell analysis. Microfluidic methods have addressed some challenges in limiting the realization of microflow cytometry, but most microfluidics-based flow cytometry techniques still rely on bulky equipment (e.g., high-precision syringe pumps and bench-top microscopes). Here, we describe a comprehensive approach that achieves high-throughput white blood cell (WBC) counting in a portable and handheld manner, thereby allowing the complete miniaturization of flow cytometry. Our approach integrates three major components: a motorized smart pipette for accurate volume metering and controllable liquid pumping, a microfluidic cell concentrator for target cell enrichment, and a miniaturized fluorescence microscope for portable flow cytometric analysis. We first validated the capability of each component by precisely metering various fluid samples and controlling flow rates in a range from 219.5 to 840.5 μL/min, achieving high sample-volume reduction via on-chip WBC enrichment, and successfully counting single WBCs flowing through a region of interrogation. We synergistically combined the three major components to create a handheld, integrated microflow cytometer and operated it with a simple protocol of drawing up a blood sample via pipetting and injecting the sample into the microfluidic concentrator by powering the motorized smart pipette. We then demonstrated the utility of the microflow cytometer as a quality control means for leukoreduced blood products, quantitatively analyzing residual WBCs (rWBCs) in blood samples present at concentrations as low as 0.1 rWBCs/μL. These portable, controllable, high-throughput, and quantitative microflow cytometric technologies provide promising ways of miniaturizing flow cytometry.

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“Organ-on-chip” specific light sheet fluorescence microscope for vascularization investigation

10.22443/rms.lsfm2020.13 ◽

2020 ◽

Author(s):

Caroline Bissardon

Keyword(s):

Light Sheet ◽

Fluorescence Microscope ◽

On Chip

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Light sheet fluorescence microscope for microfluidic chip

EPJ Web of Conferences ◽

10.1051/epjconf/202023804005 ◽

2020 ◽

Vol 238 ◽

pp. 04005

Author(s):

Caroline Bissardon ◽

Xavier Mermet ◽

Sophie Morales ◽

Frédéric Bottausci ◽

Marie Carriere ◽

...

Keyword(s):

Microfluidic Chip ◽

3D Imaging ◽

Optical Characterization ◽

Light Sheet ◽

Fluorescence Microscope ◽

Experimental Setup ◽

Simple Design ◽

Volumetric Images ◽

On Chip

We present a light sheet fluorescence microscope dedicated to image “Organ-on-chip”-like biostructures in microfluidic chip. Based on a simple design, the setup is built around the chip and its environment to allow 3D imaging inside the chip in a microfluidic laboratory. The experimental setup, its optical characterization and first volumetric images are reported.

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On-chip real-time detection and quantification of reactive oxygen species in MCF-7 cells through an in-house built fluorescence microscope

Microelectronic Engineering ◽

10.1016/j.mee.2020.111432 ◽

2020 ◽

Vol 233 ◽

pp. 111432

Author(s):

Muhsin Ali ◽

Young Su Kim ◽

Muhammad Asad Ullah Khalid ◽

Afaque Manzoor Soomro ◽

Jae-Wook Lee ◽

...

Keyword(s):

Reactive Oxygen Species ◽

Real Time ◽

Reactive Oxygen ◽

Fluorescence Microscope ◽

Oxygen Species ◽

Real Time Detection ◽

On Chip ◽

Detection And Quantification ◽

Mcf 7

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Scanning x-ray fluorescence microscopy and principal component analysis

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100133394 ◽

1992 ◽

Vol 50 (2) ◽

pp. 1752-1753

Author(s):

Brian Cross

Keyword(s):

Principal Component Analysis ◽

Fluorescence Microscopy ◽

Spatial Resolution ◽

High Speed ◽

Image Acquisition ◽

Principal Component ◽

Fluorescence Microscope ◽

Acquisition Rate ◽

X Ray ◽

Speed Up

A relatively new entry, in the field of microscopy, is the Scanning X-Ray Fluorescence Microscope (SXRFM). Using this type of instrument (e.g. Kevex Omicron X-ray Microprobe), one can obtain multiple elemental x-ray images, from the analysis of materials which show heterogeneity. The SXRFM obtains images by collimating an x-ray beam (e.g. 100 μm diameter), and then scanning the sample with a high-speed x-y stage. To speed up the image acquisition, data is acquired "on-the-fly" by slew-scanning the stage along the x-axis, like a TV or SEM scan. To reduce the overhead from "fly-back," the images can be acquired by bi-directional scanning of the x-axis. This results in very little overhead with the re-positioning of the sample stage. The image acquisition rate is dominated by the x-ray acquisition rate. Therefore, the total x-ray image acquisition rate, using the SXRFM, is very comparable to an SEM. Although the x-ray spatial resolution of the SXRFM is worse than an SEM (say 100 vs. 2 μm), there are several other advantages.

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Gut microbiome a promising target for management of respiratory diseases

Biochemical Journal ◽

10.1042/bcj20200426 ◽

2020 ◽

Vol 477 (14) ◽

pp. 2679-2696

Author(s):

Riddhi Trivedi ◽

Kalyani Barve

Keyword(s):

Respiratory Diseases ◽

New Technology ◽

Bacterial Flora ◽

Systemic Circulation ◽

The Body ◽

Lung Pathology ◽

Promising Target ◽

Current Therapies ◽

Intestinal Microbial Flora ◽

On Chip

The intestinal microbial flora has risen to be one of the important etiological factors in the development of diseases like colorectal cancer, obesity, diabetes, inflammatory bowel disease, anxiety and Parkinson's. The emergence of the association between bacterial flora and lungs led to the discovery of the gut–lung axis. Dysbiosis of several species of colonic bacteria such as Firmicutes and Bacteroidetes and transfer of these bacteria from gut to lungs via lymphatic and systemic circulation are associated with several respiratory diseases such as lung cancer, asthma, tuberculosis, cystic fibrosis, etc. Current therapies for dysbiosis include use of probiotics, prebiotics and synbiotics to restore the balance between various species of beneficial bacteria. Various approaches like nanotechnology and microencapsulation have been explored to increase the permeability and viability of probiotics in the body. The need of the day is comprehensive study of mechanisms behind dysbiosis, translocation of microbiota from gut to lung through various channels and new technology for evaluating treatment to correct this dysbiosis which in turn can be used to manage various respiratory diseases. Microfluidics and organ on chip model are emerging technologies that can satisfy these needs. This review gives an overview of colonic commensals in lung pathology and novel systems that help in alleviating symptoms of lung diseases. We have also hypothesized new models to help in understanding bacterial pathways involved in the gut–lung axis as well as act as a futuristic approach in finding treatment of respiratory diseases caused by dysbiosis.

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Clinical Update: Measuring Shoulder Joint Motion

Guides Newsletter ◽

10.1001/amaguidesnewsletters.1998.sepoct02 ◽

1998 ◽

Vol 3 (5) ◽

pp. 4-5

Author(s):

Christopher R. Brigham

Keyword(s):

External Rotation ◽

Joint Motion ◽

Fourth Edition ◽

Single Plane ◽

Reconstructive Procedures ◽

Results Of Treatment ◽

Shoulder Disorders ◽

Shoulder Motion ◽

Motion Measurements ◽

Upper Extremity Impairment

Abstract Accurate measurement of shoulder motion is critical in assessing impairment following shoulder disorders. To this end, measuring and recording joint motion are important steps in diagnosing, determining the severity and progression of a disorder, assessing the results of treatment, and evaluating impairment. Shoulder movement usually is composite rather than in a single plane, so isolating single movements is challenging. Universal goniometers with long arms are used to measure shoulder motion, and testing must be performed and recorded consistently. Passive motion may be carried out cautiously by the examiner; two measurements of the same patient by the same examiner should lie within 10° of each other. Shoulder extension and flexion are illustrated. Maximal flexion of the shoulder also includes slight external rotation and abduction, and controlling or eliminating these components during evaluation is challenging. Abduction and adduction are illustrated. Deficits in external rotation may occur in patients who have undergone reconstructive procedures with an anterior approach; deficits in internal rotation may result from issues with shoulder instability. The authors recommend recording the shoulder's range of motion measurements according to the Upper Extremity Impairment Evaluation Record in the AMA Guides to the Evaluation of Permanent Impairment, Fourth Edition.

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