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Bioengineering
定量的な光学顕微鏡:細胞生物物理の測定は、標準的な光学顕微鏡で特徴
定量的な光学顕微鏡:細胞生物物理の測定は、標準的な光学顕微鏡で特徴
JoVE Journal
Bioengineering
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JoVE Journal Bioengineering
Quantitative Optical Microscopy: Measurement of Cellular Biophysical Features with a Standard Optical Microscope

定量的な光学顕微鏡:細胞生物物理の測定は、標準的な光学顕微鏡で特徴

Full Text
16,217 Views
14:09 min
April 7, 2014

DOI: 10.3791/50988-v

Kevin G. Phillips1,2, Sandra M. Baker-Groberg1, Owen J.T. McCarty1,3

1Department of Biomedical Engineering,Oregon Health & Science University, School of Medicine, 2Department of Dermatology,Oregon Health & Science University, School of Medicine, 3Department of Cell & Developmental Biology, Division of Hematology & Medical Oncology, Knight Cancer Institute,Oregon Health & Science University, School of Medicine

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Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This article describes a method to quantify cellular mass, volume, and density using a standard optical microscope. The technique combines bright field and differential interference contrast imagery to obtain accurate measurements without the need for specimen fixation or staining.

Key Study Components

Area of Science

  • Cellular biology
  • Microscopy techniques
  • Image processing

Background

  • Quantifying physical properties of cells is essential for various biological studies.
  • Traditional methods often require fixation or staining, which can alter cellular properties.
  • This method utilizes non-invasive imaging techniques.
  • Bright field and differential interference contrast microscopy provide clear images of live cells.

Purpose of Study

  • To develop a technique for measuring cellular mass, volume, and density.
  • To demonstrate the advantages of using standard optical microscopy over fluorescence microscopy.
  • To provide a method that does not require cell fixation or staining.

Methods Used

  • Mounting cellular specimens on glass cover slips.
  • Obtaining through focus brightfield and differential interference contrast images.
  • Inputting Z stack images into MATLAB for image processing.
  • Extracting physical data from intensity measurements.

Main Results

  • Successful quantification of cellular mass, volume, and density.
  • Demonstrated the effectiveness of the imaging technique.
  • Provided a non-invasive method for analyzing live cells.
  • Highlighted the advantages over traditional methods.

Conclusions

  • This method allows for accurate measurements of cellular properties without altering the cells.
  • It opens new avenues for studying live cellular dynamics.
  • The technique can be applied to various biological research fields.

Frequently Asked Questions

What is the main advantage of this microscopy technique?
The main advantage is that it does not require cellular specimens to be fixed, permeated, or stained, allowing for live cell analysis.
What types of images are used in this method?
Bright field and differential interference contrast images are used to obtain cellular measurements.
How are the images processed?
Images are processed using MATLAB programs that extract physical data from the Z stack images.
Can this method be used for all types of cells?
Yes, this method can be applied to various types of cellular specimens grown on glass cover slips.
Is this technique suitable for quantitative analysis?
Yes, it is designed specifically for quantitative measurements of cellular mass, volume, and density.

私たちは、明視野と微分干渉コントラスト画像の組み合わせを通じて、細胞の質量、体積、密度の定量的測定を行うための標準的な光学顕微鏡の使用について説明します。

この手順の全体的な目標は、標準的な光学顕微鏡と画像処理を使用して、質量や体積など、細胞標本の基本的な物理的特徴を定量化することです。これは、まずガラスカバースリップ上に成長した細胞標本を顕微鏡スライドにマウントすることで達成されます。次に、フォーカス明視野と微分干渉コントラスト画像を通じて取得されます。

次に、イメージの各 Z スタックを個別の MATLAB 画像処理プログラムに入力し、物理データを抽出します。最終的に、細胞標本の基本的な物理的特性は、明視野コントラスト顕微鏡および微分干渉コントラスト顕微鏡下での焦点強度測定を使用して取得されます。蛍光顕微鏡法のような既存の方法に対するこの技術の主な利点は、細胞標本を固定、透過、または染色する必要がないことです。

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