酵母血小板X2アプリケーションノート

Cellometer X2 Image Cytometer
Cellometer Reagents
Optimized Analysis for Yeast and other Small Cells
Catalog #
Description
CS1-0108-5ML
Features of the Cellometer X2
User-Friendly Software and Assay Selection: Enhanced
inter-operator reproducibility, minimal training, auto-save
option
Instrument Compatibility
Size
Unit
Auto 2000, K2, X2, X1, X4, Vision CBA
5 mL
each
CS1-0109-5ML
PI (propidium iodide) Staining Solution for staining of dead nucleated cells
Auto 2000, K2, X2, X4, Vision CBA
5 mL
each
CS2-0106-5ML
AO/PI (acridine orange / propidium iodide) Staining Solution for live/dead Mammalian nucleated cells
Auto 2000, K2, X2, Vision CBA
5 mL
each
Cellometer Annexin V-FITC / PI Apoptosis Reagents
X2, K2, Vision CBA
each
K183-100-N
K183-25-N
Cellometer Caspase-3 Apoptosis Kit
X2, K2, Vision CBA
each
K188-100-N
K188-25-N
CS1-0114
CS0-0115-100ML
CS1-0116
Dual Fluorescence and Bright Field Imaging: staining of
both live and dead cells in yeast samples
AO (acridine orange) Staining Solution for staining of nucleated cells
Cellometer Caspase-8 Apoptosis Kit
X2, K2,Vision CBA
each
CSK-0112
Cellometer PI Cell Cycle Kit
X1, X2, K2, Vision CBA
each
CSK-0102
Cellometer ViaStain Kit for live/dead Yeast concentration including stainer buffer, fluorescent dye mixture
X2, K2, Vision CBA
Catalog #
Small Sample Size: Only 20 µl of sample
Broad Dynamic Range: Measurable concentration range
of 2.5 x 105 to 5 x 107 cells/mL using Nexcelom’s proprietary
de-clustering function
Description
Size
Standard chamber thickness. Packed in microscope slide boxes. Ready to use.
Case of 500 slides for 1,000 counts (10
individual boxes)
1 Case
CHT4-SD100-014
Standard chamber thickness. Packed with protective film on both sides. Remove protective film
before use.
Case of 900 slides for 1,800 counts
1 Case
CHT4-PD300-003
3x standard chamber thickness. Packed in microscope slide boxes. Ready to use.
Simply Counted
Case of 500 slides for 1,000 counts (10
1 Case
individual boxes)
See www.nexcelom.com/products for updated product selections.
Image Cytometer
é
• Verify cell morphology and counted live/dead cells
• Export cell images for presentations and publications
é
Pattern Recognition Software
• Accurately count cells in clumps
• Count irregular-shaped cells
• Eliminate debris from cell counts
• Differentiate cells based on size
é
Automated Data Management
• Pre-set assays and automated reports
• Archive sample images and auto-save results
é
Maintenance-free System
• Disposable counting chambers – no wash steps
• No required instrument maintenance
On-Line Demonstrations are completed in just 20
to 30 minutes and provide an overview of how
Cellometer works using existing images of cells
that interest you.
On-Site Demonstrations are a convenient way to
test a Cellometer system for a specific application.
An experienced Applications Specialist will arrive at
your lab for a hands-on session to test your cells and
show how Cellometer can enhance your workflow.
Technical Seminars are an excellent way to
introduce Cellometer systems to a lab group or
collaborators in different laboratories within an
organization. A trained biologist will discuss and
demonstrate the capabilities and advantages of
Cellometer image cytometry.
Call 978-327-5340 or E-mail
[email protected] today to
schedule a free demonstration
or technical seminar.
Nexcelom products are for RESEARCH USE ONLY and are not approved for diagnostic or therapeutic use.
8001236 Rev.B 5/14
© Copyright 2014 Nexcelom Bioscience LLC. All Rights Reserved.
é
Which Cellometer is Right for Me?
Cell Imaging
Unit
CHT4-PD100-003
Cellometer Cell Counters, Cell Analysis Systems & Image Cytometry
Nexcelom offers a wide range of Cellometer systems developed
and optimized for specific applications and cell types.
Many Compatible Dyes: Trypan blue, AO, PI, EB, 7AAD,
AO/PI, AO/EB, Calcein AM, CFDA-AM, Calcein AM/PI,
CFDA/PI
Learn why thousands of users,
including the top ten pharmaceutical
companies, trust Cellometer.
イメージサイトメーター
細胞カウンティング＆解析
Cellometer Counting Chamber
Fast Results: Obtain cell images, counts, size
measurements, and viability calculations in 60 seconds
Advantages of Cellometer Image Cytometer
Cellometer ® X2
Features
Automated Cell Counters
For more information, visit
www.nexcelom.com
Image Cytometers
Mini
Auto
T4
Auto
1000
Auto
2000
X4
(10x)
X1
X2
K2
Vision
CBA
Vision
CBA
(10x)
Objective Magnification
4X
4X
4X
4X
10X
10X
10X
4X
5X
10X
Cell Line
X
X
X
X
X
X
Cultured Primary Cells
X
X
X
X
X
X
Cell / Sample Type
X
Algae
X
Platelets
Low Cocnentration Cell Lines
X
X
X
X
Yeast (Clean Sample)
X
X
X
X
X
Primary cells (Messy Sample*)
X
X
X
PBMCs, Splenocytes, Stem Cells
X
X
X
X
X
X
X
X
X
X
Apoptosis (Annexin V-FITC/PI)
X
X
X
Apoptosis (Caspase Activity)
X
X
X
Autophagy (CytoID-green)
X
X
Cell Proliferation (CFSE)
X
X
X
Yeast (Messy Sample)
Hepatocytes
Adipocytes***
X
Cell-Based Assay **
X
X
X
X
X
X
X
X
X
X
X
X
YFP Transfection
X
X
RFP Transfection
X
X
Mitochondrial Potential (JC-1)
X
X
Multi-drug Resistance (ABC
Transporter)
X
X
Surface Marker Analysis
X
X
X
X
Cell Cycle (PI)
GFP Transfection
Vitality (Calcein-AM/PI)
X
X
X
Image Cytometry**
X
X
* A messy sample is a heterogeneous sample containing unwanted cell types, such as red blood cells, in addition to the cells of interest.
** FCS Express 4 license must be purchased in order to perform Cell Based Assay or Image Cytometry analysis
*** Cellometer CHT4-PD300 slides are required for cells greater than 80µm in diameter
Contact us at:
Nexcelom Bioscience
360 Merrimack Street, Building 9
Lawrence, MA 01843, USA
Email: [email protected]
Phone: 978.327.5340
Fax: 978.327.5341
・（ビール）醸造酵母
・ワイン酵母
・血小板
その他小さい細胞
セロメーター X2 イメージサイトメーター
Cellometer Reagents
酵母や小さい細胞の解析に最適です。
Catalog #
セロメーター X2の特徴
Instrument Compatibility
Size
Unit
CS1-0108-5ML
AO (acridine orange) Staining Solution for staining of nucleated cells
Description
Auto 2000, K2, X2, X1, X4, Vision CBA
5 mL
each
CS1-0109-5ML
PI (propidium iodide) Staining Solution for staining of dead nucleated cells
Auto 2000, K2, X2, X4, Vision CBA
5 mL
each
CS2-0106-5ML
AO/PI (acridine orange / propidium iodide) Staining Solution for live/dead Mammalian nucleated cells
Auto 2000, K2, X2, Vision CBA
5 mL
each
Cellometer Annexin V-FITC / PI Apoptosis Reagents
X2, K2, Vision CBA
each
K183-100-N
K183-25-N
Cellometer Caspase-3 Apoptosis Kit
X2, K2, Vision CBA
each
K188-100-N
K188-25-N
CS1-0114
CS0-0115-100ML
CS1-0116
蛍光2色と明視野でのイメージング：酵母サンプルの生
死判定が可能です。
簡便なソフトウェアと解析：再現性の強化、トレーニング
不要、自動データ保存
Cellometer Caspase-8 Apoptosis Kit
X2, K2,Vision CBA
each
CSK-0112
Cellometer PI Cell Cycle Kit
X1, X2, K2, Vision CBA
each
CSK-0102
Cellometer ViaStain Kit for live/dead Yeast concentration including stainer buffer, fluorescent dye mixture
X2, K2, Vision CBA
Cellometer ® X2
Image Cytometer
for Cell Counting & Analysis
Cellometer Counting Chamber
迅速な測定：細胞イメージの取得、カウント、サイズ
計測、生存率算出まで60秒以内
Catalog #
サンプル容量：わずか20μl
広範囲な検出可能濃度領域：ネクセローム社独自のデクラ
スター機能により、 2.5 x 105 - 5 x 107 cells/mLが検
出可能
Description
Size
Unit
CHT4-PD100-003
Standard chamber thickness. Packed in microscope slide boxes. Ready to use.
Case of 500 slides for 1,000 counts (10
individual boxes)
1 Case
CHT4-SD100-014
Standard chamber thickness. Packed with protective film on both sides. Remove protective film
before use.
Case of 900 slides for 1,800 counts
1 Case
CHT4-PD300-003
3x standard chamber thickness. Packed in microscope slide boxes. Ready to use.
Case of 500 slides for 1,000 counts (10
1 Case
individual boxes)
See www.nexcelom.com/products for updated product selections.
Cellometer Cell Counters, Cell Analysis Systems & Image Cytometry
Nexcelom offers a wide range of Cellometer systems developed
and optimized for specific applications and cell types.
様々な色素に対応：トリパンブルー、AO、PI、EB、７
AAD、AO/PI、AO/EB、Calcein
AM、Calsein
AM/PI、CFAD/PI
Brewing Yeast
Simply Counted
Wine Yeast
Image Cytometer
Platelets
セロメーターの利点
é
細胞イメージング
• 細胞の形状や生死判定の確認
• 画像をエクスポートし、プレゼン資料や論文へ使用可能
é
細胞認識ソフトウェア
• 凝集した細胞も分離して識別
• 通常と異なる形状もカウント可能
• 夾雑物をカウント対象から除外
• サイズの違いによる識別
é
自動データ管理
• 測定メニューと自動レポートを標準装備
• サンプル画像と自動データ保存
é
メンテナンスフリー
• 使い捨てのカウンティングチャンバーで洗浄不要
• 装置本体のメンテナンス不要
トップ製薬企業をはじめとするユーザーが
セロメーターを信頼する理由を
デモンストレーションで体感してください。
デモンストレーションビデオ
下記URLにて、セロメーターの使い方や測定原理の概要に
ついて知ることができます。１ビデオは数分で見ることが
できます。　※英語ビデオ
http://www.nexcelom.com/Support/Cellometer-VideoLibrary.html
オンサイトデモンストレーション
アプリケーションスペシャリストが訪問し、実際の細胞サ
ンプルを用いての測定をハンズオンでお見せします。セロ
メーターが細胞カウント作業をいかに簡単で正確なものに
するかを体感することができます。
ご用命は
トミーデジタルバイオロジー株式会社へ
電話：03－5834－0810
E-mail:[email protected]
Nexcelom products are for RESEARCH USE ONLY and are not approved for diagnostic or therapeutic use.
8001236 Rev.B 5/14
© Copyright 2014 Nexcelom Bioscience LLC. All Rights Reserved.
é
Which Cellometer is Right for Me?
Features
Automated Cell Counters
For more information, visit
www.nexcelom.com
Image Cytometers
Mini
Auto
T4
Auto
1000
Auto
2000
X4
(10x)
X1
X2
K2
Vision
CBA
Vision
CBA
(10x)
Objective Magnification
4X
4X
4X
4X
10X
10X
10X
4X
5X
10X
Cell Line
X
X
X
X
X
X
Cultured Primary Cells
X
X
X
X
X
X
Cell / Sample Type
X
Algae
X
Platelets
Low Cocnentration Cell Lines
X
X
X
X
Yeast (Clean Sample)
X
X
X
X
X
Primary cells (Messy Sample*)
X
X
X
PBMCs, Splenocytes, Stem Cells
X
X
X
X
X
X
X
X
X
X
Apoptosis (Annexin V-FITC/PI)
X
X
X
Apoptosis (Caspase Activity)
X
X
X
Autophagy (CytoID-green)
X
X
Cell Proliferation (CFSE)
X
X
X
Yeast (Messy Sample)
Hepatocytes
Adipocytes***
X
Cell-Based Assay **
X
X
X
X
X
X
X
X
X
X
X
X
YFP Transfection
X
X
RFP Transfection
X
X
Mitochondrial Potential (JC-1)
X
X
Multi-drug Resistance (ABC
Transporter)
X
X
Surface Marker Analysis
X
X
X
X
Cell Cycle (PI)
GFP Transfection
Vitality (Calcein-AM/PI)
X
X
X
Image Cytometry**
X
X
* A messy sample is a heterogeneous sample containing unwanted cell types, such as red blood cells, in addition to the cells of interest.
** FCS Express 4 license must be purchased in order to perform Cell Based Assay or Image Cytometry analysis
*** Cellometer CHT4-PD300 slides are required for cells greater than 80µm in diameter
Contact us at:
Nexcelom Bioscience
360 Merrimack Street, Building 9
Lawrence, MA 01843, USA
Email: [email protected]
Phone: 978.327.5340
Fax: 978.327.5341
and Other Small Cells
一般的に、醸造工場で使用されている酵母は非常にクリーンです。これらの酵母はセロメーターX1およびX2カウン
ト可能です。細胞濃度や生存率は明視野および蛍光測定によって算出することが可能です。
明視野での酵母の濃度測定
é
シングルセルカウント
デクラスターカウント
鎖状セルカウント
é
明視野と蛍光による酵母の濃度＆生存率測定
酵母の生存率と生育能
é
Propidium Iodide (PI)に
よる生存率測定
明視野で全細胞数を、蛍光で
死細胞をカウントし、生存率
を算出します。
é
é
Yeast Vitality
Analysis
é
Sperm Analysis
é
2色蛍光による生存率測定
酵母サンプルとアクリジンオレンジ（AO）・プロピ
ジウムアイオダイド（PI)混合液とを1：1で混合し蛍
光染色します。染色後すぐにセロメータX2で測定
し、酵母の細胞濃度と生存率を算出します。生細胞は
緑色蛍光、死細胞は赤色蛍光で検出されます。
é
Oxonolによる生存率測定
明視野で全細胞数を、蛍光で
死細胞をカウントし、生存率
を算出します。
Yeast Cell Cycle
Analysis
酵母の細胞周期解析
Performance of the Cellometer X2 Image Cytometer
8.00E+07
6.00E+07
FEATURES
Dual FL/BR
Channels
Easily Edit and
Import Assays
Concentration Dynamic Range Figur
e1
depicts the dynamic range for cell
concentration measured by Cellometer
X2. This data set was taken on a
concentration series of cultured yeasts.
y = 1.0059x
R2 = 0.99961
7.00E+07
5.00E+07
Images for
Data Verification
Cell Size
Histograms
5 – 5 x 10
7 cells/ml
Samples from 2.5 x 10
can be counted without further dilution.
4.00E+07
3.00E+07
The %CV at each concentration was
below 10%.
2.00E+07
1.00E+07
0.00E+00
0.00E+00
2.00E+07
4.00E+07
6.00E+07
Expected Value (Cells/ml)
8.00E+07
Figure 1.Table of results for cell concentration dynamic range
Viability Dynamic Range The viability dynamic range is 0 - 100% for Cellometer X2
Image Cytometer using dual fluorescence AO/PI stain.
Consistency and Repeatability
The results
indicate the accuracy of the Cellometer
X2 instrument in assessing the viability of
AVE
1.32E+07
78.1%
STDEV
7.69E+05
2.2%
yeasts using AOPI for cell viability. Yeasts
CV(%)
5.84
2.78
were tested at 24 sample replications. The
Figure 2:Table of results for cell concentration and viability using acridine orange (AO) and viability average was calculated and
propidium Iodide (PI)
plotted. The results show theeliability
r
and accuracy of the Cellometer X2
in measuring cell concentration and
viability of yeast cells.
Cellometer X2
Average Live Cell
Concentration Via Fluorescence
Viability
Cellometer Performance
ASSAYS
Performance of the Cellometer X1 Image Cytometer
Concentration (Cells/ml)
Cell Cycle Analysis Using Propidium
Iodide (PI)を用いた細胞周期解析一般的な
パン酵母をNexcelom Bioscience社のCell
Cycle Staining Kit で染色し60分インキュ
ベート後、セロメーターX2で細胞周期を解
析を行いました。左図はPI蛍光強度をヒスト
グラムで示しています。DNA量から活動期の
ポピュレーションが示されます。
Cello
from
meter
Nexcel
X2
om
Imag
Bioscie
e
nce
Cytom
eter
Viability
forYeast
Analysis by Dual
Fluorescence
Yeast
Chain Yeast
& Small
Culture Analysis
Other
Platelet Analysis
Small Cell
Small
Concentration and
Viability Analysis
Cells
Concentration (Cells/ml)
醸造工場で用いられている酵母
7.0E+07
酵素蛍光染色による生存率測定V
6.0E+07
5.0E+07
酵母サンプルとCarboxyfluorescein-AM を１：１
で混合し酵素蛍光染色した後、セロメーターX2で
生存率を算出します。明視野で全細胞を蛍光で活性
酵母細胞を検出します。
Concentration Dynamic Range Figur
e3
depicts the dynamic range for cell
concentration measured by Cellometer
X1. This data set was taken on a
concentration series of cultured yeasts.
y = 1.026x
R2 = 0.99482
4.0E+07
5 – 5 x 10
7 cells/ml can
Samples from 4 x 10
be counted without further dilution.
3.0E+07
2.0E+07
The %CV at each concentration was
below 10%.
1.0E+07
1色蛍光イメージングモード
X1
X2
○
○
○
○
2色蛍光イメージングモード
PIによる生存率
How It Works
全血中の血小板自動カウント
0.0E+00
0.0E+001.0E+072.0E+07 3.0E+07 4.0E+075.0E+076.0E+07 7.0E+07
Expected Value (Cells/ml)
é
明視野イメージングモード
↑
酵母の解析
SET UP
○
○
AOPIを用いた細胞濃度および生存率
○
Yeast AOPI
Viability
Yeast AOPI
Viability
Small Chain
east
Y Culture
Platelets
Windsor Ale
Rehydrated
Small Cell Concentration
Wine east
Y
Rehydrated PI Viability
Yeast itality
V
CFDA
AM
Sperm
Yeast Cell Cycle
○
酵母細胞周期
○
○
デクラスター認識および鎖状細胞の識別
○
○
平均直径およびサイズ分布表示
○
○
新規細胞パラメーターの作成
○
○
Viability Dynamic Range
The viability dynamic range is 0 - 100% for Cellometer X1
Image Cytometer using PI stain.
Assay
○
CFDA-AMを用いた生育能
Figure 3.Table of results for cell concentration dynamic range
Cellomete
r®
蛍光による結小板濃度の測定
Bioscience社のCalcein
血液サンプルをNexcelom
AM Vitality / Viability Kit で20分インキュベートし
て染色します。血小板と白血球が緑色蛍光を発する
ので、サイズで識別し血小板のみをカウントしま
す。
Pipette 20 µl of
Cell Sample
Insert Counting
Chamber
Select Assay &
Click Count
Get Results
Consistency and Repeatability
The results
indicate the accuracy of the Cellometer
AVE
4.07E+07
69.0%
X1 instrument in assessing the viability of
STDEV
2.28E+06
2.5%
yeasts using PI for cell viability. Yeasts
CV(%)
5.61
3.64
were tested at 24 sample replications. The
Figure 4
. Table of results for cell concentration and viability using bright-field and Propidium viability average was calculated and
iodide (PI)
plotted. The results show theeliability
r
and accuracy of the Cellometer X1
in measuring cell concentration and
viability of yeast cells.
Cellometer X1
Average Total Cell
Concentration via Bright-Field
Viability
In general, yeast strains used in the brewing industry are very clean. They are counted using Cellometer X1 and X2 Image
Cytometers. Concentration and viability are measured using Cellometer bright field and fluorescent images.
酵母や小さい細胞用セロメーター　X2
8.00E+07
Yeast Concentration Measurement by Bright Field Analysis
機能
X
X
Concentration & viability using AOPI
解析
X
Cellometer software for analysis of clumpy and
irregular-shaped cells
X
X
Mean diameter and cell size distribution
X
X
Cell type wizard for creating new cell type parameters
X
X
The %CV at each concentration was
below 10%.
2.00E+07
1.00E+07
2.00E+07
4.00E+07
6.00E+07
Expected Value (Cells/ml)
Cellometer X2
Average Live Cell
Concentration Via Fluorescence
Viability
AVE
1.32E+07
78.1%
STDEV
7.69E+05
2.2%
CV(%)
5.84
2.78
7.0E+07
Concentration (Cells/ml)
Vitality by Fluorescent Enzymatic Stain
Yeast samples are stained 1-to-1 with
Carboxyfluorescein-AM fluorescent enzymatic
stain for 45 minutes and then analyzed for vitality
using Cellometer X2. Bright field images are used
for total cell count and fluorescent images are
used to measure the active yeast cells.
操作方法
Automated Platelet Counting in Whole Blood
Consistency and Repeatability The results
indicate the accuracy of the Cellometer
X2 instrument in assessing the viability of
yeasts using AOPI for cell viability. Yeasts
were tested at 24 sample replications. The
viability average was calculated and
plotted. The results show the reliability
and accuracy of the Cellometer X2
in measuring cell concentration and
viability of yeast cells.
Concentration Dynamic Range Figure 3
depicts the dynamic range for cell
concentration measured by Cellometer
X1. This data set was taken on a
concentration series of cultured yeasts.
y = 1.026x
R2 = 0.99482
6.0E+07
5.0E+07
4.0E+07
Samples from 4 x 105 – 5 x 107 cells/ml can
be counted without further dilution.
3.0E+07
2.0E+07
The %CV at each concentration was
below 10%.
1.0E+07
0.0E+00
0.0E+00 1.0E+07 2.0E+07
3.0E+07
4.0E+07 5.0E+07 6.0E+07
7.0E+07
Expected Value (Cells/ml)
Figure 3. Table of results for cell concentration dynamic range
Cellometer®
SET UP
Viability Dynamic Range The viability dynamic range is 0 - 100% for Cellometer X1
Image Cytometer using PI stain.
Assay
Yeast AOPI Viability
Fluorescence-Based Platelet Concentration
Measurement A blood sample stained using the
Calcein AM Vitality / Viability Kit from Nexcelom
Bioscience is incubated for 20 minutes. Both
platelets and white blood cells produce green
fluorescence. Cell size gating is applied to
exclusively count platelets.
8.00E+07
Performance of the Cellometer X1 Image Cytometer
Yeast AOPI Viability
Small Chain Yeast Culture
Platelets
Windsor Ale Rehydrated
Small Cell Concentration
Wine Yeast Rehydrated PI Viability
Yeast Vitality CFDA AM
Sperm
Yeast Cell Cycle
X
X
3.00E+07
Cellometer Performance
X
Yeast cell cycle
Samples from 2.5 x 105 – 5 x 107 cells/ml
can be counted without further dilution.
4.00E+07
Figure 2: Table of results for cell concentration and viability using acridine orange (AO) and
propidium Iodide (PI)
X
Vitality using CFDA-AM
5.00E+07
Viability Dynamic Range The viability dynamic range is 0 - 100% for Cellometer X2
Image Cytometer using dual fluorescence AO/PI stain.
X
X
é
X
6.00E+07
Figure 1. Table of results for cell concentration dynamic range
é
Viability using PI
酵母の細胞周期
解析
X2
X
Dual fluorescence imaging mode
精子解析
é
Single fluorescence imaging mode
酵母の生育能解析
é
é
Bright field imaging mode
小さい細胞の濃度・
生死判定
é
Cell Cycle Analysis Using Propidium Iodide
(PI) Standard baker’s yeast stained with
the Cell Cycle Staining Kit from Nexcelom
Bioscience are incubated for 60 minutes
before using Cellometer X2 to analyze
the cell cycle. The plot shows the yeast
population that is actively dividing. Their
higher DNA content is measured using PI.
血小板の解析
é
Yeast Cell Cycle Analysis
é
Viability by Dual-Fluorescence Yeast samples are
stained 1-to-1 with a mixture of acridine orange
(AO) and propidium iodide (PI) dual-fluorescence
stain. Yeast concentration and viability are
obtained immediately after staining using
Cellometer X2. Live yeast cells fluoresce green
and dead cells fluoresce red.
小さい鎖状酵母の
培養解析
Concentration Dynamic Range Figure 1
depicts the dynamic range for cell
concentration measured by Cellometer
X2. This data set was taken on a
concentration series of cultured yeasts.
y = 1.0059x
R2 = 0.99961
7.00E+07
0.00E+00
0.00E+00
２色蛍光で酵母の
生死判定
é
Viability Measurement by
Oxonol Bright field images
are used to obtain total cell
count, while fluorescent
images are used to count
dead cells.
X1
細胞サイズを
ヒストグラム
Yeast Viability and Vitality
Viability Measurement
Using Propidium Iodide (PI)
Bright field images are used
to obtain total cell count,
while fluorescent images
are used to count dead
cells.
Cellometer Yeast Analysis
画像でデータを
検証
Chain-Forming Cell Count
é
Yeast Concentration & Viability Measurement by Bright Field
& Fluorescence
解析プログラム
を簡単に編集・
インポート
é
De-clustering of Yeast Cells
2色蛍光/BR
é
é
Single Cell Count
Performance of the Cellometer X2 Image Cytometer
Concentration (Cells/ml)
Yeast Used in Brewing Industry
サンプルを20μl
充填
スライドチャン
バーを本体へ挿入
Assayを選択し、
カウントボタンを
クリック
Cellometer X1
Average Total Cell
Concentration via Bright-Field
Viability
AVE
4.07E+07
69.0%
STDEV
2.28E+06
2.5%
CV(%)
5.61
3.64
Figure 4. Table of results for cell concentration and viability using bright-field and Propidium
iodide (PI)
結果
Consistency and Repeatability The results
indicate the accuracy of the Cellometer
X1 instrument in assessing the viability of
yeasts using PI for cell viability. Yeasts
were tested at 24 sample replications. The
viability average was calculated and
plotted. The results show the reliability
and accuracy of the Cellometer X1
in measuring cell concentration and
viability of yeast cells.
In general, yeast strains used in the brewing industry are very clean. They are counted using Cellometer X1 and X2 Image
Cytometers. Concentration and viability are measured using Cellometer bright field and fluorescent images.
Cellometer X2 Image Cytometer for Yeast & Other Small Cells
from Nexcelom Bioscience
Yeast Concentration Measurement by Bright Field Analysis
é
é
Images for
Data Verification
Cell Size
Histograms
Small Chain Yeast
Culture Analysis
é
Platelet Analysis
é
Small Cell
Concentration and
Viability Analysis
é
Yeast Vitality
Analysis
é
Sperm Analysis
é
Viability by Dual-Fluorescence Yeast samples are
stained 1-to-1 with a mixture of acridine orange
(AO) and propidium iodide (PI) dual-fluorescence
stain. Yeast concentration and viability are
obtained immediately after staining using
Cellometer X2. Live yeast cells fluoresce green
and dead cells fluoresce red.
Yeast Viability
Analysis by Dual
Fluorescence
é
Viability Measurement
Using Propidium Iodide (PI)
Bright field images are used
to obtain total cell count,
while fluorescent images
are used to count dead
cells.
y = 1.0059x
R2 = 0.99961
7.00E+07
6.00E+07
Yeast Cell Cycle
Analysis
Figure 1 にセロメーターX2で酵母の希
釈系列を測定したグラフを示します。
4.00E+07
2.5 x 105 – 5 x 107 cells/ml の範囲で
は、希釈することなくカウントすることが
可能です。
3.00E+07
各濃度のCV（％）は10％以下でした。
5.00E+07
2.00E+07
1.00E+07
0.00E+00
0.00E+00
Yeast Viability and Vitality
Viability Measurement by
Oxonol Bright field images
are used to obtain total cell
count, while fluorescent
images are used to count
dead cells.
Easily Edit and
Import Assays
Chain-Forming Cell Count
é
Yeast Concentration & Viability Measurement by Bright Field
& Fluorescence
é
De-clustering of Yeast Cells
Dual FL/BR
Channels
細胞濃度の測定範囲
8.00E+07
FEATURES
é
Single Cell Count
セロメータX2の性能評価
Concentration (Cells/ml)
Yeast Used in Brewing Industry
2.00E+07
4.00E+07
6.00E+07
Expected Value (Cells/ml)
8.00E+07
Figure 1. 細胞濃度の測定可能範囲
生死判定の範囲
AO/PIを用いた２色蛍光染色によって、セロメーターX2で0 - 100% の範囲で生死判定できます。
セロメーターX2
蛍光による細胞濃度測定
整合性と再現性
AOPIを用いた生死判定の正確性をFigure 2
に示します。24回繰り返し測定し、生存率の
平均を算出しました。
この結果から、酵母の細胞濃度および生存率
をセロメーターX2で測定した時の信頼性と再
現性が示されました。
生存率
AVE
1.32E+07
78.1%
STDEV
7.69E+05
2.2%
CV(%)
5.84
2.78
Figure 2: AO/PI染色による細胞濃度と生存率の測定結果
セローメーターの性能
Yeast Cell Cycle Analysis
Single fluorescence imaging mode
7.0E+07
Vitality by Fluorescent Enzymatic Stain
Yeast samples are stained 1-to-1 with
Carboxyfluorescein-AM fluorescent enzymatic
stain for 45 minutes and then analyzed for vitality
using Cellometer X2. Bright field images are used
for total cell count and fluorescent images are
used to measure the active yeast cells.
How It Works
X1
X
X
X
Automated Platelet Counting in Whole Blood
Concentration & viability using AOPI
SET UP
X
Yeast AOPI Viability
Cellometer software for analysis of clumpy and
irregular-shaped cells
X
X
Mean diameter and cell size distribution
X
X
Cell type wizard for creating new cell type parameters
X
X
Fluorescence-Based Platelet Concentration
Measurement A blood sample stained using the
Calcein AM Vitality / Viability Kit from Nexcelom
Bioscience is incubated for 20 minutes. Both
platelets and white blood cells produce green
fluorescence. Cell size gating is applied to
exclusively count platelets.
3.0E+07
2.0E+07
各濃度の %CVは10％以下でした。
1.0E+07
3.0E+07
4.0E+07 5.0E+07 6.0E+07
7.0E+07
生死判定の範囲
PIを用いた蛍光染色によって、セロメーターX１で0 - 100% の範囲で生死判定できます。
Assay
Yeast AOPI Viability
Small Chain Yeast Culture
Platelets
Windsor Ale Rehydrated
Small Cell Concentration
Wine Yeast Rehydrated PI Viability
Yeast Vitality CFDA AM
Sperm
Yeast Cell Cycle
X
X
4 x 105 – 5 x 107 cells/mlの濃度範囲では
サンプルを希釈することなく測定することが
可能です。
4.0E+07
(Cells/ml) Figure 3. 細胞濃度の測定可能範囲
Cellometer®
X
Yeast cell cycle
5.0E+07
Expected Value
X
Vitality using CFDA-AM
y = 1.026x
R2 = 0.99482
6.0E+07
0.0E+00 1.0E+07 2.0E+07
X
X
細胞濃度の測定範囲
Figure 3ではセロメータX1で酵母希釈系
列の細胞濃度を測定した時の濃度範囲を示
しています。
0.0E+00
X2
X
Dual fluorescence imaging mode
Viability using PI
セロメーター　X1の性能
é
Bright field imaging mode
é
Cellometer Yeast Analysis
ASSAY S
Concentration (Cells/ml)
Cell Cycle Analysis Using Propidium Iodide
(PI) Standard baker’s yeast stained with
the Cell Cycle Staining Kit from Nexcelom
Bioscience are incubated for 60 minutes
before using Cellometer X2 to analyze
the cell cycle. The plot shows the yeast
population that is actively dividing. Their
higher DNA content is measured using PI.
Pipette 20 µl of
Cell Sample
Insert Counting
Chamber
Select Assay &
Click Count
セロメーター　X1
明視野での細胞濃度測定
AVE
4.07E+07
69.0%
STDEV
2.28E+06
2.5%
CV(%)
5.61
3.64
Figure 4. 明視野とPropidium iodide (PI)による細胞濃度と生存率の測定結果
Get Results
生存率
整合性と再現性
Figure 4ではセロメーターX1を用い、PIで
酵母の細胞濃度と生存率を24回測定した時の
正確性を示しています。

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