Calibration for single multi-mode fiber digital scanning microscopy imaging system

2015 ◽  
Author(s):  
Zhe Yin ◽  
Guodong Liu ◽  
Bingguo Liu ◽  
Yu Gan ◽  
Zhitao Zhuang ◽  
...  
Keyword(s):  
Imaging System ◽  
Scanning Microscopy ◽  
Microscopy Imaging ◽  
Digital Scanning ◽  
Multi Mode

Multi-mode light microscopy of microtubule assembly dynamics and chromosome movement in vivo and In vitro

1996 ◽  
Vol 54 ◽  
pp. 730-731
Author(s):  
E. D. Salmon ◽  
J. C. Waters ◽  
C. Waterman-Storer
Keyword(s):  
Imaging System ◽  
Ccd Camera ◽  
Transmitted Light ◽  
Microtubule Assembly ◽  
Live Cells ◽  
Optical Efficiency ◽  
Multiple Band ◽  
Multi Mode

We have developed a multi-mode digital imaging system which acquires images with a cooled CCD camera (Figure 1). A multiple band pass dichromatic mirror and robotically controlled filter wheels provide wavelength selection for epi-fluorescence. Shutters select illumination either by epi-fluorescence or by transmitted light for phase contrast or DIC. Many of our experiments involve investigations of spindle assembly dynamics and chromosome movements in live cells or unfixed reconstituted preparations in vitro in which photodamage and phototoxicity are major concerns. As a consequence, a major factor in the design was optical efficiency: achieving the highest image quality with the least number of illumination photons. This principle applies to both epi-fluorescence and transmitted light imaging modes. In living cells and extracts, microtubules are visualized using X-rhodamine labeled tubulin. Photoactivation of C2CF-fluorescein labeled tubulin is used to locally mark microtubules in studies of microtubule dynamics and translocation. Chromosomes are labeled with DAPI or Hoechst DNA intercalating dyes.

Design and Experiment of Parallel Detection of Fluorescence Confocal Microscopy Imaging System

2015 ◽  
Vol 44 (8) ◽  
pp. 811002
Author(s):  
王青青 WANG Qing-qing ◽  
郑继红 ZHENG Ji-hong ◽  
桂坤 GUI Kun ◽  
王康妮 WANG Kang-ni ◽  
李道萍 LI Dao-ping ◽  
...  
Keyword(s):  
Confocal Microscopy ◽  
Imaging System ◽  
Microscopy Imaging ◽  
Parallel Detection ◽  
Fluorescence Confocal Microscopy ◽  
Confocal Microscopy Imaging

Multi-Mode Light Microscopy of Microtubule and Endoplasmic Reticulum Dynamics in Migrating Newt Epithelial Cells.

1997 ◽  
Vol 3 (S2) ◽  
pp. 211-212
Author(s):  
C. M. Waterman-Storer ◽  
E. D. Salmon
Keyword(s):  
Digital Imaging ◽  
Imaging System ◽  
Ccd Camera ◽  
Diaphragm Position ◽  
Multiple Band ◽  
Fluorescence Light ◽  
Band Pass ◽  
Cooled Ccd ◽  
System 1 ◽  
Multi Mode

We have developed a multi-mode digital imaging system (1-3) which acquires images with a 12 bit cooled CCD camera. A multiple band pass dichromatic mirror and robotically controlled excitation filter wheels provide rapid wavelength selection for epi-fluorescence with DAPI, fluorescein or GFP and X-rhodamine fluorophores while maintaining image registration on the cooled CCD detector. Shutters select illumination either by epi-fluorescence or by transmitted light for phase contrast or DIC. A robotically controlled emission filter wheel in front of the CCD camera inserts an analyzer in the light path for DIC imaging. To maximize fluorescence light intensity, the analyzer is removed and an optical flat of equivalent optical thickness is inserted for fluorescence imaging. A slider is inserted at the field diaphragm position of the fluorescence epi-illuminator to provide in-focus slit and spot targets for 360 nm photoactivation of “caged” fluorophores. The microscope system is robotically controlled and image acquisition and analysis is performed using MetaMorph™ digital imaging software.

Computed aided classification of circulating tumor cell and CK+ subclasses.

2012 ◽  
Vol 30 (30_suppl) ◽  
pp. 54-54
Author(s):  
Oleg Gusyatin ◽  
David Tims ◽  
Aladin Milutinovic ◽  
Chunsheng Jiang
Keyword(s):  
Prostate Cancer ◽  
Cancer Patients ◽  
Classification System ◽  
Imaging System ◽  
High Sensitivity ◽  
Automatic Identification ◽  
Cell Capture ◽  
Microscopy Imaging ◽  
Artifact Rejection ◽  
Normal Controls

54 Background: Fluorescence microscopy imaging system (OnQView, On-Q-ity, Waltham, MA) in combination with advanced cell capture techniques (OnQChip, On-Q-ity, Waltham, MA) provides necessary sensitivity to detect circulating tumor cells (CTCs) in a blood sample. The detection process involves automatic identification of CTC candidates from the collected imagery followed by CTC subclass identification. Subclass identification process is manual and usually leads to increased sample processing time. Methods: We have developed a fully automated CTC detection and classification system allowing for substantial increase in throughput while maintaining high sensitivity and specificity. Detection is accomplished by a robust segmentation technique. A set of 25 image-based features is automatically computed for each detected candidate. Features include texture measurements, morphology measurements, multichannel intensity and contextual characteristics. All CTC subclasses as well as artifact classes are manually labeled and verified by trained imaging technologists.A hierarchy of Multi-Layer Perceptron Neural Network (MLPNN) classifiers is then trained and used to identify and reject artifacts and to identify CTC subclasses. Results: A total of 27 prostate cancer patients and 33 normal controls with two 3.75ml blood samples per patient were used to validate techniques. Probability of successful artifact rejection was achieved to be 0.78 and probabilities of subsequent successful CTC subclass identification ranged between 0.79 and 0.98 (intact CTCs = 95%; irregular CTCs = 98%; fragmented CTCs = 82%). Conclusions: A fully automated CTC detection and classification system was developed. Testing was conducted with 27 prostate cancer patients and 33 normal controls to yield an artifact rejection probability of 0.78 and CTC subclass identification probabilities of 0.79 to 0.98.

scholarly journals Multiview Deconvolution Two-photon Laser Scanning Microscopy

2020 ◽  
Author(s):  
Dimitrios Kapsokalyvas ◽  
Rodrigo Rosas ◽  
Rob Janssen ◽  
Jo Vanoevelen ◽  
Martin Strauch ◽  
...  
Keyword(s):  
Second Harmonic ◽  
Light Sheet ◽  
Laser Scanning Microscopy ◽  
Three Dimensions ◽  
Scanning Microscopy ◽  
Fluorescence Signal ◽  
Microscopy Imaging ◽  
Two Photon ◽  
Light Sheet Microscopy ◽  
3D Image Reconstruction

Abstract Imaging in three dimensions is necessary for thick tissues and small organisms. This is possible with tomographic optical microscopy techniques such as confocal, two-photon and light sheet microscopy. All these techniques suffer from anisotropic resolution and limited penetration depth. In the past, Multiview microscopy - imaging the sample from different angles followed by 3D image reconstruction - was developed to address this issue for light sheet microscopy based on fluorescence signal. In this study we applied this methodology to accomplish Multiview imaging with two-photon microscopy based on fluorescence and additionally second harmonic signal from myosin and collagen. It was shown that isotropic resolution was achieved, the entirety of the sample was visualized, and interference artifacts were suppressed allowing clear visualization of collagen fibrils and myofibrils. This method can be applied to any scanning microscopy technique without microscope modifications. It can be used for imaging tissue and whole mount small organisms such as heart tissue, and zebrafish larva in 3D, label-free or stained, with at least 3-fold axial resolution improvement which can be significant for the accurate quantification of small 3D structures.

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