Horiba Scientific DeltaMyc J810018 User manual
DeltaMyc
Fluorescence Lifetime
Imaging Microscope System
Operation Manual
Part number J810018
DeltaMyc Operation Manual J810018 (7 Oct 2014)
ii
Copyright © 2013, 2014 by HORIBA Instruments Incorporated. All rights reserved. No
part of this work may be reproduced, stored, in a retrieval system, or transmitted in any
form by any means, including electronic or mechanical, photocopying and recording,
without prior written permission from HORIBA Instruments Incorporated. Requests for
permission should be requested in writing. Windows®is a trademark of Microsoft Cor-
poration.
Information in this manual is subject to change without notice, and does not represent a
commitment on the part of the vendor.
October 2014
Part Number J810018
DeltaMyc Operation Manual J810018 (7 Oct 2014)
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Table of Contents
0: Introduction ........................................................................................... 0-1
About the DeltaMyc..........................................................................................................................0-1
Chapter overview .............................................................................................................................0-2
Disclaimer.........................................................................................................................................0-3
Safety summary ...............................................................................................................................0-5
Risks of ultraviolet exposure ............................................................................................................0-7
Additional risks of intense light sources...........................................................................................0-9
Laser light.......................................................................................................................................0-11
CE compliance statement..............................................................................................................0-12
1: Anatomy of the DeltaMyc ............................................................................ 1-1
Introduction.......................................................................................................................................1-1
Block diagram...................................................................................................................................1-2
DeltaMyc-Inverted............................................................................................................................1-3
DeltaMyc-Upright .............................................................................................................................1-4
2: Requirements & Installation ........................................................................ 2-1
Safety-training requirements............................................................................................................2-1
Surface requirements.......................................................................................................................2-2
Environmental requirements............................................................................................................2-3
Electrical requirements.....................................................................................................................2-4
Unpacking and Installation...............................................................................................................2-5
3: System Operation...................................................................................... 3-1
Introduction.......................................................................................................................................3-1
Controlling the hardware..................................................................................................................3-3
Single-point measurement.............................................................................................................3-11
Lifetime mapping............................................................................................................................3-16
Data-analysis using the DAS6 Foundation-level package.............................................................3-25
4: Maintenance & Troubleshooting .................................................................... 4-1
Maintenance.....................................................................................................................................4-1
Troubleshooting ...............................................................................................................................4-2
Further assistance… ........................................................................................................................4-6
5: USB Port Configuration ............................................................................... 5-1
6: Technical Specifications ............................................................................ 6-1
Introduction.......................................................................................................................................6-1
Microscope.......................................................................................................................................6-1
Sample compartment.......................................................................................................................6-1
Excitation sources............................................................................................................................6-1
Motorized stage................................................................................................................................6-1
TCSPC electronics...........................................................................................................................6-2
Detector............................................................................................................................................6-2
Filters................................................................................................................................................6-2
Software...........................................................................................................................................6-2
Dimensions.......................................................................................................................................6-2
Electrical...........................................................................................................................................6-2
Environment.....................................................................................................................................6-3
Minimum host-computer requirements ............................................................................................6-3
7: Index .................................................................................................... 7-1
DeltaMyc Operation Manual J810018 (7 Oct 2014)
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DeltaMyc Operation Manual J810018 (7 Oct 2014) Introduction
0-1
Note:
Keep this and the other reference manuals near the system.
0: Introduction
About the DeltaMyc
The DeltaMyc is a time-correlated single-photon counting (TCSPC) fluorescence life-
time system designed for the study of microscopic samples. It is the ideal system to
measure lifetimes ranging from 100 ps to 10 µs, on micron-scale features. Its confocal
and imaging capabilities makes it the ultimate tool for fluorescence microscopy inves-
tigation, with high spatial resolution and high timing sensitivity.
The system consists of the following components:
Microscope with motorized x,y stage and CCD camera
Confocal pinhole adapter
DeltaHub TCSPC timing electronics
Direct-coupled light source from the DeltaDiode series
DeltaDiode controller
PPD detector (near-IR, up to 900 nm, options available)
host computer with pre-installed data-acquisition cards and software: DataStation
measurement and control software; DAS decay-analysis software (Foundation-level
package)
The timing electronics are controlled through the intuitive DataStation measurement
control software application. Decay data measured using DataStation can be analyzed
using the DAS decay-analysis software package.
This manual explains how to operate and maintain a DeltaMyc microscope. The manu-
al also describes measurements and tests essential to obtain accurate data. Please refer
to the DataStation and DAS operation manuals for more information about how to use
the software. An in-depth description of the hardware items can also be found in their
respective manuals; we advise you to familiarize yourself with them. This operation
manual is primarily concerned with the installation, and providing an overview of the
DeltaMyc system operation, with measurement examples.
DeltaMyc Operation Manual J810018 (7 Oct 2014) Introduction
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Chapter overview
0: Introduction
Safety information
1: Anatomy of the DeltaMyc
Overall interconnection of the various parts of the DeltaMyc
2: Requirements & Installation
Power and environmental requirements; select the best spot
for the instrument and set-up.
3: System Operation
Operation of the fluorometer system, and calibration in-
structions.
4: Maintenance & Trouble-
shooting
Routine cleaning, and what to do should there be a problem.
5: USB Port Configuration
How the USB ports are configured.
6: Technical Specification
7: Index
DeltaMyc Operation Manual J810018 (7 Oct 2014) Introduction
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Disclaimer
By setting up or starting to use any HORIBA Instruments Incorporated product, you are
accepting the following terms:
You are responsible for understanding the information contained in this document. You
should not rely on this information as absolute or all-encompassing; there may be local
issues (in your environment) not addressed in this document that you may need to ad-
dress, and there may be issues or procedures discussed that may not apply to your situa-
tion.
If you do not follow the instructions or procedures contained in this document, you are
responsible for yourself and your actions and all resulting consequences. If you rely on
the information contained in this document, you are responsible for:
Adhering to safety procedures
Following all precautions
Referring to additional safety documentation, such as Material Safety Data Sheets
(MSDS), when advised
As a condition of purchase, you agree to use safe operating procedures in the use of all
products supplied by HORIBA Instruments Incorporated, including those specified in
the MSDS provided with any chemicals and all warning and cautionary notices, and to
use all safety devices and guards when operating equipment. You agree to indemnify
and hold HORIBA Instruments Incorporated harmless from any liability or obligation
arising from your use or misuse of any such products, including, without limitation, to
persons injured directly or indirectly in connection with your use or operation of the
products. The foregoing indemnification shall in no event be deemed to have expanded
HORIBA Instruments Incorporated’s liability for the products.
HORIBA Instruments Incorporated products are not intended for any general cosmetic,
drug, food, or household application, but may be used for analytical measurements or
research in these fields. A condition of HORIBA Instruments Incorporated’s ac-
ceptance of a purchase order is that only qualified individuals, trained and familiar with
procedures suitable for the products ordered, will handle them. Training and mainte-
nance procedures may be purchased from HORIBA Instruments Incorporated at an ad-
ditional cost. HORIBA Instruments Incorporated cannot be held responsible for actions
your employer or contractor may take without proper training.
Due to HORIBA Instruments Incorporated’s efforts to continuously improve our prod-
ucts, all specifications, dimensions, internal workings, and operating procedures are
subject to change without notice. All specifications and measurements are approximate,
based on a standard configuration; results may vary with the application and environ-
ment. Any software manufactured by HORIBA Instruments Incorporated is also under
constant development and subject to change without notice.
Any warranties and remedies with respect to our products are limited to those provided
in writing as to a particular product. In no event shall HORIBA Instruments Incorpo-
DeltaMyc Operation Manual J810018 (7 Oct 2014) Introduction
0-4
rated be held liable for any special, incidental, indirect or consequential damages of any
kind, or any damages whatsoever resulting from loss of use, loss of data, or loss of
profits, arising out of or in connection with our products or the use or possession there-
of. HORIBA Instruments Incorporated is also in no event liable for damages on any
theory of liability arising out of, or in connection with, the use or performance of our
hardware or software, regardless of whether you have been advised of the possibility of
damage.
DeltaMyc Operation Manual J810018 (7 Oct 2014) Introduction
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Safety summary
The following general safety precautions must be observed during all phases of opera-
tion of this instrument. Failure to comply with these precautions or with specific warn-
ings elsewhere in this manual violates safety standards of design, manufacture and in-
tended use of instrument. HORIBA Instruments Incorporated assumes no liability for
the customer’s failure to comply with these requirements. Certain symbols are used
throughout the text for special conditions when operating the instruments:
A WARNING notice denotes a hazard. It calls at-
tention to an operating procedure, practice, or sim-
ilar that, if incorrectly performed or adhered to,
could result in personal injury or death. Do not
proceed beyond a WARNING notice until the in-
dicated conditions are fully understood and met.
HORIBA Instruments Incorporated is not responsi-
ble for damage arising out of improper use of the
equipment.
A CAUTION notice denotes a hazard. It calls at-
tention to an operating procedure, practice, or sim-
ilar that, if incorrectly performed or adhered to,
could result in damage to the product. Do not pro-
ceed beyond a CAUTION notice until the indicat-
ed conditions are fully understood and met.
HORIBA Instruments Incorporated is not responsi-
ble for damage arising out of improper use of the
equipment.
Laser light! Wear protective goggles, full-face
shield, skin-protection clothing, and light-blocking
gloves. Do not look directly at the beam and al-
ways observe appropriate safety procedures.
Ultraviolet light! Wear protective goggles, full-
face shield, skin-protection clothing, and UV-
blocking gloves. Do not stare into light.
Intense ultraviolet, visible, or infrared light! Wear
light-protective goggles, full-face shield, skin-
protection clothing, and light-blocking gloves. Do
not stare into light.
Caution:
Caution:
Warning:
Caution:
Warning:
DeltaMyc Operation Manual J810018 (7 Oct 2014) Introduction
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Risk of electric shock! This symbol warns the user
that un-insulated voltage within the unit may have
sufficient magnitude to cause electric shock.
Danger to fingers! This symbol warns the user that
the equipment is heavy, and can crush or injure the
hand if precautions are not taken.
This symbol cautions the user that excessive hu-
midity, if present, can damage certain equipment.
Hot! This symbol warns the user that hot equip-
ment may be present, and could create a risk of
fire or burns.
Read this manual before using or servicing the in-
strument.
Wear protective gloves.
Wear appropriate safety goggles to protect the
eyes.
Wear an appropriate face-shield to protect the
face.
General information is given concerning operation
of the equipment.
Note:
Caution:
Caution:
Caution:
Caution:
DeltaMyc Operation Manual J810018 (7 Oct 2014) Introduction
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Risks of ultraviolet exposure
Do not aim the UV light at anyone.
Do not look directly into the light.
Always wear protective goggles, full-face shield and skin protection clothing and
gloves when using the light source.
Light is subdivided into visible light, ranging from 400 nm (violet) to 700 nm (red);
longer infrared, “above red” or > 700 nm, also called heat; and shorter ultraviolet
radiation (UVR), “below violet” or < 400 nm. UVR is further subdivided into UV-
A or near-UV (320–400 nm), also called black (invisible) light; UV-B or mid-UV
(290–320 nm), which is more skin penetrating; and UV-C or far-UV (< 290 nm).
Health effects of exposure to UV light are familiar to anyone who has had sunburn.
However, the UV light level around some UV equipment greatly exceeds the level
found in nature. Acute (short-term) effects include redness or ulceration of the skin.
At high levels of exposure, these burns can be serious. For chronic exposures, there
is also a cumulative risk of harm. This risk depends upon the amount of exposure
during your lifetime. The long-term risks for large cumulative exposure include
premature aging of the skin, wrinkles and, most seriously, skin cancer and cataract.
Damage to vision is likely following exposure to high-intensity UV radiation. In
adults, more than 99% of UV radiation is absorbed by the anterior structures of the
eye. UVR can contribute to the development of age-related cataract, pterygium,
photodermatitis, and cancer of the skin around the eye. It may also contribute to
age-related macular degeneration. Like the skin, the covering of the eye or the cor-
nea, is epithelial tissue. The danger to the eye is enhanced by the fact that light can
enter from all angles around the eye and not only in the direction of vision. This is
especially true while working in a dark environment, as the pupil is wide open. The
lens can also be damaged, but because the cornea acts as a filter, the chances are re-
Caution:
This instrument is used in conjunction with ultra-
violet light. Exposure to these radiations, even reflected
or diffused, can result in serious, and sometimes irre-
versible, eye and skin injuries.
Overexposure to ultraviolet rays threatens human health
by causing:
Immediate painful sunburn
Skin cancer
Eye damage
Immune-system suppression
Premature aging
DeltaMyc Operation Manual J810018 (7 Oct 2014) Introduction
0-8
duced. This should not lessen the concern over lens damage however, because cata-
racts are the direct result of lens damage.
Burns to the eyes are usually more painful and serious than a burn to the skin. Make
sure your eye protection is appropriate for this work. NORMAL EYEGLASSES OR
CONTACTS OFFER VERY LIMITED PROTECTION!
Training
For the use of UV sources, new users must be trained by another member of the labora-
tory who, in the opinion of the member of staff in charge of the department, is suffi-
ciently competent to give instruction on the correct procedure. Newly trained users
should be overseen for some time by a competent person.
Caution:
UV exposures are not immediately felt. The us-
er may not realize the hazard until it is too late and the
damage is done.
DeltaMyc Operation Manual J810018 (7 Oct 2014) Introduction
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Warning:
Bright light
sources are danger-
ous. Please read the
following precautions.
Additional risks of intense light sources
Among the dangers associated with bright light
sources are:
Burns caused by contact with a hot lamp.
Fire ignited by hot lamp.
Interaction of other nearby chemicals with intense ultraviolet, visible, or infrared
radiation.
Damage caused to apparatus placed close to the lamp.
Explosion or mechanical failure of the lamp.
Visible radiation
Any very bright visible light source will cause a human aversion response: we either
blink or turn our head away. Although we may see a retinal afterimage (which can last
for several minutes), the aversion response time (about 0.25 seconds) normally protects
our vision. This aversion response should be trusted and obeyed. NEVER STARE AT
ANY BRIGHT LIGHT-SOURCE FOR AN EXTENDED PERIOD. Overriding the
aversion response by forcing yourself to look at a bright light-source may result in per-
manent injury to the retina. This type of injury can occur during a single prolonged ex-
posure. Excessive exposure to visible light can result in skin and eye damage.
Visible light sources that are not bright enough to cause retinal burns are not necessari-
ly safe to view for an extended period. In fact, any sufficiently bright visible light
source viewed for an extended period will eventually cause degradation of both night
and color vision. Appropriate protective filters are needed for any light source that
causes viewing discomfort when viewed for an extended period of time. For these rea-
sons, prolonged viewing of bright light sources should be limited by the use of appro-
priate filters.
The blue-light wavelengths (400–500 nm) present a unique hazard to the retina by
causing photochemical effects similar to those found in UV-radiation exposure.
Infrared radiation
Infrared (or heat) radiation is defined as having a wavelength between 780 nm and 1
mm. Specific biological effectiveness “bands” have been defined by the CIE (Commis-
sion Internationale de l’Éclairage or International Commission on Illumination) as fol-
lows:
• IR-A (near IR) (780–1400 nm)
• IR-B (mid IR) (1400–3000 nm)
• IR-C (far IR) (3000 nm–1 mm)
DeltaMyc Operation Manual J810018 (7 Oct 2014) Introduction
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The skin and eyes absorb infrared radiation (IR) as heat. Workers normally notice ex-
cessive exposure through heat sensation and pain. Infrared radiation in the IR-A that
enters the human eye will reach (and can be focused upon) the sensitive cells of the ret-
ina. For high irradiance sources in the IR-A, the retina is the part of the eye that is at
risk. For sources in the IR-B and IR-C, both the skin and the cornea may be at risk from
“flash burns.” In addition, the heat deposited in the cornea may be conducted to the lens
of the eye. This heating of the lens is believed to be the cause of so called “glassblow-
ers’ ” cataracts because the heat transfer may cause clouding of the lens.
Retinal IR Hazards (780 to 1400 nm): possible retinal lesions from acute high irra-
diance exposures to small dimension sources.
Lens IR Hazards (1400 to 1900 nm): possible cataract induction from chronic lower
irradiance exposures.
Corneal IR Hazards (1900 nm to 1 mm): possible flashburns from acute high irradi-
ance exposures.
Who is likely to be injured? The user and anyone exposed to the radiation or lamp
shards as a result of faulty procedures. Injuries may be slight to severe.
DeltaMyc Operation Manual J810018 (7 Oct 2014) Introduction
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Warning:
Laser light can damage your eyes. Never
stare into a laser beam.
Laser light
Practice care when inserting samples, especially highly reflective samples under the ob-
jectives, for they can reflect or scatter the excitation light. Also see safety notices relat-
ing to light sources.
Some DeltaDiode excitation sources generate laser light. Do not look directly at the
beam and always observe appropriate safety procedures. Always refer to the infor-
mation provided on the label attached to the DeltaDiode. If in doubt please contact your
organization’s laser safety officer.
DeltaMyc Operation Manual J810018 (7 Oct 2014) Introduction
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DeltaMyc Operation Manual J810018 (7 Oct 2014) Anatomy of the DeltaMyc
1-1
1: Anatomy of the DeltaMyc
Introduction
The DeltaMyc is made up of a large number of devices and hardware. Therefore this
chapter is designed to help you understand how all the devices are connected and inter-
act with each other.
DeltaMyc Operation Manual J810018 (7 Oct 2014) Anatomy of the DeltaMyc
1-2
Block diagram
Following is a block diagram showing how the major sections of the DeltaMyc are inter-
connected.
Table of contents
