IBiDI Microfluidics

Immunofluorescence

Chemotaxis Assays

Angiogenesis Assays

Live Cell Imaging

Migration Assays

Flow Assays

Imaging Chambers

Cells & Reagents

For more information on our products, and to download our full catalog, please visit us at: www. .com

2

ibid

i’s I

mag

ing

Ch

amb

ers

for

Eve

ry L

ab

ibidi’s Wide Range of Imaging Chambers

Find the perfect imaging chamber for your own application!

• High-resolution microscopy through a coverslip-like bottom

• Excellent cell growth on tissue culture treated surface (ibiTreat)

• Proven mechanical and chemical stability

• Broad range of sizes and coatings

µ-Slide 2 well | 4 well | 8 wellChambered coverslips combining optimal cell culture and high-resolution microscopy

µ-Slide VI 0.4 | µ-Slide VI 0.1

A 6 channel µ-Slide for immunofluorescence and parallel flow assays

µ-Dish FamilyPetri dishes with a coverslip-like bottom for high end microscopy

NEW: µ-Slide 2 well Ph+ | µ-Slide 4 well Ph+ Excellent optical quality in phase contrast without meniscus formation

Ordering Information:

ibiTreat – the standard surface for live cell imaging

100 xCoverslip for High Resolution

80286µ-Slide 2 well, ibiTreat



80296µ-Slide 2 well Ph+, ibiTreat


81156µ-Dish 35 mm, high, ibiTreat

81166µ-Dish 35 mm, high Grid -500, ibiTreat

81136µ-Dish 50 mm, low, ibiTreat

81191µ-Dish 35 mm, high ESS, uncoated elastic surface

80606µ-Slide VI 0.4, ibiTreat

3

µ-Plate Angiogenesis 96 wellA µ-Plate 96 well used to investigate angiogenesis in high throughput tube formation assays

µ-Plate FamilyBlack, multi-well plates (24, 96, 384 wells) with flat and clear bottom for high throughput applications in cell-based assays

Plates

sticky-Slide Chemotaxis 3D A chemotaxis bottomless slide used in 2D and 3D applications with a self- adhesive underside designed to allow self-assemblage of the slide setup

µ-Slide Chemotaxis 2D

Used to investigate chemotaxis of slow migrating adherent cells on 2D surfaces

µ-Slide Chemotaxis 3D

Used to investigate chemotaxis of adherent cells, and non-adherent cells in gel matrices – also available as a sticky-Slide

µ-Slide III 3in1

A µ-Slide for switchable chemo- tactic gradients that merges three separated liquids into one channel

Ch

emo

taxis Slid

es

µ-Slide IA combination of a cell culture channel and a coverslip for imaging inside a channel

µ-Slide I Luer FamilyChannel slides with different heights, volumes, and coatings specially suited for flow applications – also available as a sticky-Slide

µ-Slide III 0.1

A three channel µ-Slide for flow assays and immunofluorescence that uses a minimal volume of reagents

µ-Slide y-shapedA flow-through µ-Slide for bifurcation studies and blood vessel simulation

Flow

Ch

amb

ers

sticky-Slide VI 0.4 A 6-channel bottomless µ-Slide with a self-adhesive underside to which your own substrates can be mounted

Culture-Insert FamilySilicone inserts with a defined cell- free gap, suitable for wound healing, migration assays, 2D invasion assays, and co-cultivation of cells

µ-Chamber 12 wellA removable silicone cultivation chamber for cell culture and immunofluorescence staining

micro-Insert 4 well FamilyA four well silicone insert for live cell imaging of both adherent and suspension cells

Rem

ovable Cham

bers

sticky-Slide 8 well An 8 well bottomless µ-Slide with a self-adhesive underside to which your own substrates can be mounted

µ-Slide 2 x 9 wellA µ-Slide suitable for co-cultivation assays in combination with microscopy

µ-Slide AngiogenesisA µ-Slide used to investigate angiogenesis in tube formation assays. Also perfect for 3D cell culture and immunofluorescence staining

µ-Slide 18 well – flatA µ-Slide with 18 wells for use in matrix tests

Op

en S

lides

Order a free sample at

www.ibidi.com

to test ibidi’s µ-Slides, µ-Dishes, and µ-Plates with your experiments.

ibidi FREE SAMPLES

4

Cells & Reagents

Fuse-It Membrane Fusion – Transiently Incorporate Proteins and Particles into Cells

• Fast results within minutes

• 80 - 100 % efficiency

• Independent of cell type, cell density, and developmental status

• Applicable for dyes, proteins, lipids, beads, etc.

LifeAct™ – The Actin Marker for Visualization of F-Actin in Living Cells

• Brilliant visualization of F-actin in live cell imaging

• Unrestricted actin functionality and dynamics

• Available as a plasmid, adenoviral or lenti-viral vector, and HT-1080 stable cell line

Cell Culture Reagents

• ibidi Freezing Medium: Serum-free freezing of cells with extremely high recovery rates

• ibidi Mounting Medium: Conserves your fluorescent samples for months

• ibidi Anti-Evaporation Oil: Prevents medium evaporation in cell culture

Fuse-ItLiposomal

Carrier

MembraneFusion

Cytoplasm

Nucleus

Torpedo Transfection Reagents – Optimized for Use with ibidi µ-Slides and µ-Dishes

• Superb biocompatibility with low cytotoxicity for use in live cell imaging

• Transfection efficiency optimized for microscopic assays

12

6

39


60101Plasmid p CMV LifeAct- TagGFP2

60121Adenovirus rAV CMV-LifeAct- TagGFP2

60141Lentivirus rLV Ubi-LifeAct-TagGFP2

40101HT-1080 LifeAct-TagGFP2 cells

60610Torpedo DNA Transfection Reagent

60620Torpedo siRNA Transfection Reagent

60200Fuse-It green

60220Fuse-It P

80021ibidi Freezing Medium Classic

50051ibidi Anti-Evaporation Oil

5

100 x

Immunofluorescence

• Fast and easy sample preparation

• Homogeneous cell distribution

• Low amount of cells and reagents needed

• High-resolution imaging


ibidi’s All-in-One Chambers reduce the number of experimental steps needed in immunofluorescence assays.

Human umbilical vein endothelial cells (HUVEC) cultured under flow conditions in µ-Slide I 0.4 Luer

Cell line Madin-Darby canine kidney (MDCK) cultured in µ-Slide VI 0.4

The removable µ-Chamber 12 well allows for standard, immunofluorescence assays using standard glass coverslip techniques.

Application Examples:

Order a free sample at

www.ibidi.com

to test ibidi’s µ-Slides, µ-Dishes, and µ-Plates with your experiments.

ibidi FREE SAMPLES

View detailed handling steps in the movie,

Immunofluorescence Using the µ-Slide VI (MV 18),

on www.ibidi.com.

ibidi MOVIE

1. Seed cells

1. Cultivation 2. Fixation 3. Staining 4. Imaging

2. Incubate 3. Remove the chamber

4. Stain and mount

5. Store







80406µ-Dish 35 mm, high micro-Insert 4 well

81201µ-Chamber 12 well

50001ibidi Mounting Medium

6

ibidi Heating & Incubation Systems for Live Cell Imaging

• Excellent illumination of the sample without condensation

• Full incubator conditions on the microscope for time lapse video microscopy

• For use with all slides, dishes, and multi-well plates

0 h

2 h

6 h

Investigation of Tube Formation in Angiogenesis Research

2D and 3D Chemotaxis Experiments

Migration of a dendritic cell in a chemotactical gradient.

HUVEC cells on Matrigel™ in a µ-Slide Angiogenesis.

Migration and Proliferation Assays

Closure of cell-free gap in an ibidi Culture-Insert.

0 h

12 h

24 h

Experimental Examples:

The independently controlled, heated glass lid of the ibidi Heating & Incubation System solves the problem of conden-sation developing in live cell imaging.

Heated plate

Heated lidibidi solution:

40 °C

37 °C

Heated plate

Non-heated lid

34 °C

37 °C

Heated plate

Heated lidibidi solution:

40 °C

37 °C

Heated plate

Non-heated lid

34 °C

37 °C

Phase contrast without condensation Phase contrast with condensation


10918ibidi Heating System, Universal Fit

10926ibidi Heating System, Multi-Well Plates

74001ibidi OPAL Optical O2 Measurement System

11920ibidi Gas Incubation System for CO2

89626µ-Plate 96 well, ibiTreat

80326µ-Slide Chemotaxis 3D, ibiTreat

81506µ-Slide Angio-genesis, ibiTreat

81176µ-Dish 35 mm, high Culture-Insert, ibiTreat

7

The Heating Solution for All Microscopes

• Easy to install and use

• Compatible for all inverted microscope models

• Accurate and precise heating

There are two different heating system options you can order, depending on your needs. ibidi also offers an additional gas incubation unit to complete your heating system set-up.

ibidi Heating System, Universal Fit

Compatible for all inverted microscopes.

View example application movies on www.ibidi.com/support/movies

ibidi MOVIE

ibid

i Heatin

g S

ystem,

Un

iversal Fit

ibidi Heating System, Multi-Well Plates

Heating System for the Nikon TI-S-E and TI-S-ER Motorized Stage which holds multi-well plates (for low magnification only).

ibidi Temperature Controller

Heated Plate with Heated Glass Bottom

Heated Glass Lid, Multi-Well Plate

Multi-Well Plate

ibid

i Heatin

g S

ystem,

Mu

lti-Well P

lates

ibidi Temperature Controller

Heated Plate in multi-well format

Heated Glass Lid, Universal Fit

Heating Insert for ibidi and non-ibidi slide formats

ibidi Gas Incubation System for CO2 and O2

Suitable for various experimental conditions (e.g., pH or hypoxia)

Gas In

cub

ation

Systemibidi

Gas MixerHumidifying Column

+ + +

+ + +

+

NEW: Measure Oxygen Directly in Cell Cultures and Tissues Using the ibidi OPAL Optical O2 Measurement System

ibidi TIP

O2 = 10.4 % O2 = 8.7 %

8

Chemotaxis Assays

• Complete solution, from sample preparation to quantitative data analysis

• Chemotaxis measurement in real-time• Stable gradients for 48 hours in 2D or 3D chemotaxis assays• Reliable and user-independent data

Migration of a dendritic cell in a chemotactical gradient

Chemotaxis and cell polarization of a stable LifeAct expressing HT-1080 cell line

Video Microscopy

Cell Tracking

Sample Preparation

0 min 10 min 20 min

Quantitative and Statistical Analysis

10-12

10-8

10-4

10 0

p-v

alu

e(R

ayle

igh

-Tes

t)

+/- -/- +/+

0.3

0.2

0.1

0.0

FMI

+/- -/- +/+

Visual Analysis

Working Principle:

Applications:

For in vitro chemotaxis measurements of cells that are attached to a 2D surface or embedded in a 3D gel matrix.

Cell Tracking and Data Analysis: WimTaxis – Chemotaxis Image Analysis

A web-based, quantitative image analysis of chemotaxis assays with automated tracking of label-free cell lines and bacteria in phase contrast


1. Seed cells with gel matrix

2. Fill with chemoattractant-free medium

3. Fill with chemoattractant

C0

C100

1 mm

Cells in 3D matrix

70 µm

C100 C0

Examples:

- Cell polarization- Molecular aspects of chemotaxis- Collective cell migration- Single cell chemotaxis



80328sticky-Slide Chemotaxis 3D

80316µ-Slide III 3in1, ibiTreat


30003WimTaxis Image Analysis

50201Collagen Type I

9

Wound Healing & Migration Assays

• Complete solution for wound healing experiments, requiring only a few steps from sample preparation to image analysis

• Reproducible experiments owing to: a defined 500 µm cell free gap, no leakage during cultivation, and no material being left behind after the insert’s removal

1. Prepare the Culture-Insert on a flat, clean surface

2. Seed cells and wait for cell attachment

3. Remove Culture-Insert

4. Fill with medium

5. Microscopy of cell-free gap

Working Principle:

Applications:

- Wound healing

- Migration

Data Acquisition and Analysis Using WimScratch

A web-based, automated image analysis of wound healing and migration assays

Influence of inhibitors or enhancers on wound healing

2D invasion assay (Data provided by C. Matern and K.D. Nnetu, University Leipzig, Germany)

600x103

500

400

300

Cel

l Co

vere

d A

rea

[µm

²]

2520151050

Time [h]

Add drug

Normal

Enhancer

Inhibitor

- 2D invasion

- Co-cultivation of cells

Microscopy

0 h

12 h

24 h

Image Analysis


20

25

30

35

15

5

10

0

Control

Inhib

itor 1

Inhib

itor 2

Enhance

r 1

Enhance

r 2

Cel

l Fro

nt V

eloc

ity[µ

m/h

]

600x10³

500

400

300

Cel

l Cov

ered

Are

a[µ

m²]

2520151050

Time [h]


Sample Preparation

81176µ-Dish 35 mm, high Culture-Insert, ibiTreat

80241Culture-Insert 24

8020925 Culture-Inserts for Self-Insertion


11920ibidi Gas Incubation System for CO2

30002WimScratch Image Analysis

81156µ-Dish 35 mm, high, ibiTreat

10

Microscopy

Image Analysis


Sample PreparationAngiogenesis Assays

• Complete solution for tube formation experiments, requiring only a few steps from sample preparation to image analysis

• Brilliant visualization without meniscus formation, and with all cells in one focal plane

• Cost-effective experiments, requiring only 10 µl gel per well

• Objective and reproducible analysis within minutes using WimTube image analysis

ibidi’s „Well-in-a-Well“ Compared to a Standard Well

10x

1)

2)

10x

1)

2)

µ-Plate Angiogenesis 96 well

1) Planar air-liquid interface: good phase contrast all over the observation area

2) Planar gel surface: all cells are in one optical plane

Volume of Matrigel: 10 µl

Standard Well

1) Meniscus on air-liquid interface: poor phase contrast in most of the observation area

2) Mensicus on the gel surface: not possible to focus on all cells simultaneously

Volume of Matrigel: 100 µl

Applications:

- Tube formation assays

- Sprouting assays

- 3D cell culture

- Immunofluorescence

Experimental Example:

2500

2000

1500

1000

500Tu

be

Len

gth

[µ

m/m

m]

]

86420

Time [h]

Inhibitor

Normal

Influence of inhibitors on tube formation over time. Please note the time depen- dence of a tube formation and take images at identical time points.

Data Acquisition and Analysis: WimTube

A web-based, automated image analysis of tube formation assays


0 h

2 h

6 h

WimTube Key MetricsCovered Area % 36Total Tube Length [px] 12916Total Branching Points 58Total Loops 24Total Nets 1

Tube CharacteristicsTotal Tube Length [px] 12916Total Tubes 98Mean Tube Length [px] 132Std. Dev. Tube Length [px] 82

Loop CharacteristicsTotal Loops 24Mean Loop Area [px] 23809Std. Dev. Loop Area [px] 27614Mean Loop Perimeter [px] 614Std. Dev. Loop Perimeter [px] 427

10x10³

8

6

4

Tube

leng

th[µ

m/m

m²]

1086420

Time [h]

8000

6000

4000

2000

0

Tube

Len

gth

[µm

*]*

Leng

ht in

µm

/ S

tand

ard

Are

a (=

1 cm

²)

Contro

l

1 ng/

ml

10 n

g/m

l

100 n

g/m

l

1 µg/

ml

10 µ

g/m

l

Concentration

81506µ-Slide Angio-genesis, ibiTreat

89646µ-Plate Angio- genesis 96 well, ibiTreat

30001WimTube Image Analysis

30004WimSprout Image Analysis




50051ibidi Anti-Evaporation Oil

11

Flow Assays

• Ideal simulation of various physiological conditions – continuous unidirectional, oscillating, and pulsatile flow

• Suitable for long-term live cell imaging

• Minimal amount of medium and supplement

• Various types of flow chambers available

Live Cell Imaging and Immunofluorescence for Analyzing Shear Stress Response:

Human umbilical vein endothelial cells (HUVEC) cultured under flow conditions in µ-Slide I 0.4 Luer

Cell Seeding Into Channel Slides

Flow Conditioning of Adherent Cells

24 h

Staining and Image Acquisition

Types of Assays:

Cell Culture Under Shear Stress

Rolling and Adhesion Assays

Stop Flow Experiments

Applications:

Conditioning of endothelial cells under flow

Rolling and adhesion of suspended cells to protein surfaces

Defined liquid exchange



10902ibidi Pump System

80176µ-Slide I 0.4 Luer, ibiTreat



10962Perfusion Set

10802Elbow Luer Connector (and more flow accessories)

10991µ-Galaxy Cell Culture Incubator


ibidi GmbHAm Klopferspitz 1982152 Martinsried (München)Germany

Toll free within Germany: Phone: 0800 / 00 11 11 28 Fax: 0800 / 00 11 11 29

International calls:Phone: +49 89 / 520 46 17 - 0Fax: +49 89 / 520 46 17 - 59

E-Mail: [email protected]

ibidi, LLC419 Venture CourtVerona, WI 53593 USA

Toll free within the US:Phone: +1 800 372 61 02Fax: +1 608 845 32 71

E-Mail: [email protected]

Certified ISO 9001, ISO 13485

All ibidi products are for research use only! Errors and omissions excepted.

© ibidi GmbH, V 2 / 2014 / 09

For more information on our products, and to download our full catalog, please visit us at:

www. .com

Documents

IBiDI Microfluidics