Cellvizio lab In Service

www.cellviziolab.com

IN VIVO MOLECULAR IMAGINGWITH CELLULAR RESOLUTION

IN SERVICEPRESENTATION

http://www.cellviziolab.com



Disclaimer

• This presentation is not a substitute for the Instructions For Use provided with Cellvizio, including but not limited to the Cellvizio User Guide, the Quick Guide, the Fast System Setup and the Probe Disinfection User Guide.

• Always refer to the last revisions of the Instructions For Use provided with Cellvizio.




Agenda

System description Before the Procedure Probe Handling

After the Procedure Troubleshooting Contact Us




System Description: pCLE

ProFlex™ Microprobes•Designed for different applications•High Resolution: up to 1.4 µm•Thin Diameter: down to 300 µm

Laser Scanning Unit•Confocal Microscope•488 or 660 nm excitation beam•High Sensitivity(APD)•Handy, turn-key, easy-to-use

ImageCell™ Software•Real-Time image processing•Quantification features•Framerate up to 200 fps•LSU control

Foot Pedal•ImageCell™ remote control




High technology•Simultaneous injection and collection•Patented technology•High sensitivity

Each fiber acts as a point source and a point detector or pinhole

«»

CONFOCALFLUORESCENCEDETECTIONIMAGER

System Description: pCLE




OS+

OS++

Optical SectioningOS+

Working DistanceWD

WD+ WD++

Confocal Microprobes TURN TOOPTICALBIOPSY

Tailored by Application




Model ApplicationsTip

Diameter (mm)

Lateral Resolution

(µm)

Optical Sectioning

(µm)

Working Distance

(µm)

Max Field Of view

(µm)

Deep brain imaging, designed for permanent implantation on freely moving mice

Part of the NeuroPak™ solution

0,350,47 3,3 15 0 325

Brain, deep brain in mice, other organs at depth if low invasiveness is mandatory 0,3 3,3 15 0 300

Surface Imaging

Brain, deep brain in rats, other organs at depth if low invasiveness is mandatory 0,65 3,3 15 0 600

General applicability, can be used to check fluorescence sensitivity in most targets 1,5 3,3 15 0 600

Vessels, angiogenesis, cell fate, cell morphology, utility depends on cell layer thickness and invasiveness

2,6 1,4 10 60 240

Hi-ResImaging

Vessels, angiogenesis, cell fate, cell morphology, utility depends on cell layer thickness and invasiveness

4,2 1,4 10 30 240

ImagingVessels, angiogenesis, cell fate, cell morphology, utility depends on cell layer thickness and invasiveness

4,2 1,4 10 100 240

Blood flow through the vessel (without penetration) image deeper cell layers of tumor, organ or tissue

1,8 3,5 70100/170

at488/660

600

Depth Imaging

Cavities, eye 0,94 3,5 30

50/70at

488/660 325

SSeries

MSeries

S-300*

S-0650*

S-1500

UltraMiniO

MiniO/30

MiniO/100

Z-1800

Mini-Z

ZSeries

CerboFlex™CerboFlex™ J

> Made of thousands fiber-optics

Confocal Microprobes



www.cellviziolab.com8

Preparing Cellvizio®

Connect the M

icroprobe

Lock the co

nnector Clockwise

Cellvizio®

Imaging

Microprobe Warm

-Up

15 minutes Calib

ration

Using th

e Quantik

it™

Launch ImageCell™

Select a Storage Dire

ctory

Clean the M

icroprobe

Using th

e Cletop®-S

Turn ON th

e System




• Turn On computer• Turn On LSU- On/Off switch at the rear of the LSU- press On/Off on front face- look for 1st green light

Log into Cellvizio accountlogin: Cellvizio

password: cellvizio2004

TURN ON THE SYSTEM




login : Administratorpassword : admin

• Click on the ImageCell™ Software icon on the desktop or in the Dock

• Log into the Cellvizio user account of your choice (by default: administrator)

ImageCell™ 3.8.3

LAUNCH THE SOFTWARE




Hold the Cletop-S system in one hand with the blue ratchet facing upwards.

Press down the blue ratchet to uncover the cleaning ribbon.

Hold the ratchet down while positioning the Confocal Microprobe’s connector (black proximal part) on the cleaning ribbon.

Perform a ¼ turn (90°) of the connector while keeping it in contact with the cleaning ribbon.

Slide the connector optical interface along the cleaning ribbon while maintaining contact.

1

3

2

5

4

Do not use the Cletop-S cleaning system to clean

the probe’s distal end

Clean the Confocal Microprobe’s connector




1. Remove yellow cap from LSU. Then turn the LSU connector wheel counterclockwise until it blocks.

2. Hold the Microprobe with the flat side and the arrow on the label facing upwards.

3. Insert the Confocal Miniprobe™ connector in the female portion on the LSU until it blocks.

4. Hold the connector in place, and turn the wheel clockwise until it clicks. Keep turning until a 2nd click.

5. Turn the connector wheel for another 45° (1/8 turn) clockwise after the 2nd click.

6. Second green light appears on LSU

15

2

Always check that the probe has been cleaned and is not damaged

Connect the Microprobe to the LSU

1

23

4


• Once the Confocal Microprobe is connected, click on start laser to initiate the warm-up.

• The warm-up should take at least 15 minutes to be fully completed.

• Follow the calibration procedure as described on the screen, using the corresponding vials of the Quantikit

• The calibration procedure should be completed before each procedure

MAKE SURE QUANTIKIT HAS NOT EXPIRED

Warm-up and calibration


1 Calibration: enables real time image processing

2 Warm-up: reduces fibers’ fluorescence background

Optimized image quality

Raw image Processed image

Why a warm-up and a calibration?



www.cellviziolab.comAcquisition

LUT

Toolbox

Toolbox


START/STOP

LASER START/STOP

RECORDING

«Freeze» LUT

Imaging control with the foot pedal



www.cellviziolab.comBEST IMAGES = clean probe + good calibration

Calibration done too long ago

Probe has moved in the connector

RECALIBRATE

Bad image with “snow effect”

How to get THE BEST image quality

1 2OR

MAKE SURE QUANTIKITS HAVE NOT EXPIRED



Troubleshooting

If Microprobe is dirty

Probe handling: Soft contact with tissue

Clean the distal tip using the cleaning vial + QTip and check if image is clear

If not, unplug the probe and clean the proximal part with Cletop-S

CLEAN DIRTY

CENTERING FAILED

Unplug the probe and plug it again




After the Procedure

Turn off the laser Disconnect the probe Put yellow cap on LSU

Clean and rinse the probe

Dry it

Carefully fold the

microprobe

1 2 3

4

Prepared by H. Gharbi | © Mauna Kea Technologies

After the procedure• Catch the animal and anesthetize it• Place it under the stereotaxic frame• Attach the CerboFlex body to the stereotaxic holder• Carefully unlock the CerboFlex screw on the implant’s guide• Slightly remove the CerboFlex from the implant• Clean the CerboFlex tip by keeping the Laser ON for onscreen check• Put the protection cap into the implant’s cannula• Carefully remove the CerboFlex microprobe from the holder• Fold the probe as shown

22

Note 1 : NeuroPak contents & How to pack the Cerboflex

Disconnect the microprobe by turning the Cellvizio’s connector clockwise until the click. Then, release and remove the CerboFlex.

1


Put the distal and proximal protection back over the tip of the CerboFlex.


2

Note 1 : N

euroPak contents & How

to pack the Cerboflex

Curve the fiber into a circle as shown. The diameter should fit the CerboFlex print in the storage box.

3

CAUTION: Do NOT twist or fold the optical fiber bundle of the CerboFlex

Prepared by H. Gharbi | © Mauna Kea Technologies

After the procedure• Catch the animal and anesthetize it• Place it under the stereotaxic frame• Attach the CerboFlex body to the stereotaxic holder• Carefully unlock the CerboFlex screw on the implant’s guide• Slightly remove the CerboFlex from the implant• Clean the CerboFlex tip by keeping the Laser ON for onscreen check• Put the protection cap into the implant’s cannula• Carefully remove the CerboFlex microprobe from the holder• Fold the probe as shown

22


Disconnect the microprobe by turning the Cellvizio’s connector clockwise until the click. Then, release and remove the CerboFlex.

1


Put the distal and proximal protection back over the tip of the CerboFlex.


2

Note 1 : N

euroPak contents & How

to pack the Cerboflex

Curve the fiber into a circle as shown. The diameter should fit the CerboFlex print in the storage box.

3

CAUTION: Do NOT twist or fold the optical fiber bundle of the CerboFlex

5




Post-processing with IC Viewer

Double-click on the corresponding movie icon to view it

Icon Description

• Superimpose a micrometric scale on the image

• Measure a distance* from a fixed point of reference

• Take a snapshot and export it

• Select some frames of the entire sequence and export it

Find your recorder sequences in dataImageCell

Process the sequence

* measurement values are indicative and should be considered as such

IC Viewer

1

2

3




Quick and easy exportsSelect frames1

Choose export format2

3. Export3

From IC Viewer

From ImageCell

Exporting Cellvizio sequences




Always use clean and disinfected probes

Do not use undue force on probes during procedure

Do not cover the top of the LSU nor put the screen on it

Use only recommended cleaning and disinfection procedures.

Read instruction guides completely

! !

General Safety Information




Laser labeling and status

Do not stare with naked eyes or optical instruments at the laser beam emitted by the Confocal Miniprobe™

Laser class: Class 2M (resp. Class II) according to IEC 60825-1:2007 (resp. 21 CFR Part 1040.10 and 1040.11)

For detailed laser safety information, please refer to the Cellvizio User Guide

! !

LSU

Back panelFront panelThird green light

Laser Safety Information

ImageCell™



Email: [email protected]

Website: www.cellviziolab.com

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Cellvizio lab In Service