PHỤ KIỆN KÍNH HIỂN VI

Giới thiệu chung
Phụ kiện & Tùy chọn
Tài liệu kỹ thuật
Ứng dụng

	Sản phẩm	Ưu điểm	Ứng dụng chính
Light Sources	ScopeLite 200	Low cost broad band fluorescence illuminator	General Fluorescence GFP, FITC, DAPI, fluorescein, Rhodamine, Fluo-3/4 Ca⁺⁺
DeltaRAM™ X	Tunable from 250 to 650 nm Switches between any λ pair in 2 ms	Intracellular ratiometric fluorescence Fura-2 {Ca⁺⁺], FRET, BCECF [pH}
Tunable PowerArc™ Micro-Illuminator	Low cost tunable illuminator from 250 to 1,100 nm	General Fluorescence Scanning Illuminator

Sản phẩm

Ưu điểm

Ứng dụng chính

Thông tin khác

Light Sources

ScopeLite 200

Low cost broad band fluorescence illuminator

General Fluorescence GFP, FITC, DAPI, fluorescein, Rhodamine,
Fluo-3/4 Ca⁺⁺

DeltaRAM™ X

Tunable from 250 to 650 nm Switches between any λ pair in 2 ms

Intracellular ratiometric fluorescence Fura-2 {Ca⁺⁺], FRET, BCECF [pH}

Tunable PowerArc™
Micro-Illuminator

Low cost tunable illuminator from 250 to 1,100 nm

General Fluorescence
Scanning Illuminator

	Sản phẩm	Ưu điểm	Ứng dụng chính
Imaging Cameras	EFiS	UV EMCCD	Fluorescence, Live Cell Imaging
AToR	VIS EMCCD	Fluorescence, Live Cell Imaging
iRiS	High Resolution sCMOS	Fluorescence, Live Cell Imaging

Sản phẩm

Ưu điểm

Ứng dụng chính

Thông tin khác

Imaging Cameras

EFiS

UV EMCCD

Fluorescence, Live Cell Imaging

AToR

VIS EMCCD

Fluorescence, Live Cell Imaging

iRiS

High Resolution sCMOS

Fluorescence, Live Cell Imaging

	Sản phẩm	Ưu điểm	Ứng dụng chính	Thông tin khác
PMT Point Detectors	PMT Photometers, Single, Dual and Triple Detection	Millisecond kinetics Ultimate sensitivity & dynamic range	Quantitative fluorescence Indo-1, FRET Electrophysiology

Sản phẩm

Ưu điểm

Ứng dụng chính

Thông tin khác

PMT Point Detectors

PMT Photometers, Single, Dual and Triple Detection

Millisecond kinetics Ultimate sensitivity & dynamic range

Quantitative fluorescence
Indo-1, FRET Electrophysiology

	Sản phẩm	Ưu điểm	Ứng dụng chính	Thông tin khác
Micro-Spectrometers	MicOS	Low cost luminescence system, multi-wavelength excitation, broad spectral range	Photoluminescence, Electroluminescence, Raman, Photoreflectance

Sản phẩm

Ưu điểm

Ứng dụng chính

Thông tin khác

Micro-Spectrometers

MicOS

Low cost luminescence system, multi-wavelength excitation, broad spectral range

Photoluminescence, Electroluminescence, Raman, Photoreflectance

Nhà sản xuất: HORIBA Scientific

Các phụ kiện kính hiển vi của HORIBA có sẵn dưới dạng các thiết bị độc lập, tuy nhiên, nếu khách hàng đang tìm kiếm một hệ thống hoàn chỉnh với phần mềm điều khiển và các khả năng phân tích, vui lòng xem xét các hệ thống của PTI

Hệ thống quang kế PTI RatioMaster™ và phần mềm
Hệ thống chụp ảnh PTI EasyRatioPro và phần mềm

Nếu khách hàng đang tìm kiếm hệ thống kính hiển vi huỳnh quang, vui lòng xem xét sản phẩm MicOS.

Mặc dù các phụ kiện kính hiển vi của HORIBA có thể được sử dụng cho nhiều ứng dụng khác nhau nhưng chủ yếu vẫn dùng cho các ứng dụng kính hiển vi huỳnh quang định tính trong sinh học tế bào.

Intracellular Ca++
FRET Microscopy
Electrophysiology
GFP, FITC, Fluorescein, Rhodamine

Các bài ứng dụng

Intracellular Calcium

: Brian Research, 921(1-2): 1-11, 2001.
Courtesy of Dr. G. Brewer

Fura-2/AM loaded neurons
Illuminator: PTI DeltaRAM
Camera: Sensys CCD
Software: PTI ImageMaster™
Imaging: Typical Fura-2-fluorescence ratio imaging for intracellular Ca²⁺ in hippocampal neurons from old rats before NMDA (A) and after NMDA (B), scale values in nM

Single HEK293 Cell Ca2+ Imaging

: PNAS 101 (35), 13062-13067. 2004
Courtesy of Dr. S. Chen

HEK293 cell loaded with Fura-2.
Illuminator: PTI DeltaRAM
Software: PTI ImageMaster™
In the picture:(A) Single-cell fluorescent Ca²⁺ images in the presence (Upper) or absence (Lower) of 0.3 mM caffeine at various [Ca²⁺]o (0-1.0 mM).
(B) Fura-2 ratios of representative RyR2(wt) cells in the absence (green trace) and presence (blue trace) of 0.3 mM caffeine and a HEK293 parental cell expressing no RyR2 (pink trace)

Dopamine-induced Ca2+ Transients in Rat MSN

: J. Biol. Chem.,279 (40), 42082-42094, 2004.
Courtesy of Dr. I. Bezprozvanny

EGFP transfected rat MSN cultures loaded with Fura2/AM
Illuminator: PTI DeltaRAM
Camera: PTI IC-300
Software: PTI ImageMaster™ Pro
Fura-2 340/380 nm ratios in rat MSN before (-1 min) and after (0–28 min) application of 400 µM dopamine

Intracellular Calcium in Human Neuronal Cell Cultures

: J. of Neuroimmu, 98(2): 185-200, 1999.
Courtesy of Dr. H. Gendelman

Fura-2AM loaded human neurons
Illuminator: PTI DeltaRAM
Camera: Photometrics CCCD
Software: PTI ImageMaster™
Imaging: Fura-2 imaging in SDF-1 treated (B) and control (A) neuronal cells. SDF-1 activate intracellular calcium

Mitochondria [Ca2+]m in Single Cells

: J. of Neuroimmu, 98(2): 185-200, 1999.
Courtesy of Dr. H. Gendelman
EMBO, 18(1): 96-108,
Courtesy of Dr. Hajnoczky

Fura2FF-loaded single permeabilized RBL cells
PTI DeltaRAM illuminator
Photometrics PXL CCCD camera

Measurement of Extracellular Near-membrane [Ca2+]

: J Cell Sci, 2003, 116(pt 8): 1527-38.
Courtesy of Dr. Hofer, A. M

Fura-C18-loaded HEK CaR cells
Illuminator: PTI DeltaRAM
Camera: PTI IC-100
Images a–c: ratio images taken at different time points
Image d shows fluorescence at 340 nm excitation (510 nm emission) of the same cells

Mechanical Stimulation Increases Ca2+ Waves

: Am J Physiol Lung Cell Mol Physiol 280: L221-L228, 2001.
Courtesy of Dr. S. Boitano

Fura-2 AM loaded rat alveolar epithelial cells (AECs)
Illuminator: PTI DeltaRAM Camera: PTI ICCD camera
Software: PTI ImageMaster™
Images: A-D: Mechanical stimulation resulted in a Ca²⁺ wave that averaged slightly over 4 cells
E–H: in the presence of the gap junction-inhibiting peptide Gap 27, [Ca²⁺]i increase restricted to the stimulated cell
I–L: Apyrase did not significantly reduce Ca²⁺ wave propagation
Arrow: Cell that was briefly stimulated with a glass micropipette
White lines, cell borders
Color bar, approximate [Ca²⁺]i

Mechanical Wounding Increases Ca2+ Waves

: Am J Physiol Lung Cell Mol Physiol 280: L221-L228, 2001.<br/ >Courtesy of Dr. S. Boitano

Fura-2 AM loaded rat alveolar epithelial cells (AECs)
Illuminator: PTI DeltaRAM
Camera: PTI ICCD camera
Software: PTI ImageMaster™
Images: A–D: mechanical wound-induced Ca²⁺ waves
E–H: gap junction inhibitor does not affect this Ca²⁺ waves
I–L: apyrase restricts this Ca²⁺ waves
Arrow: Cell that was mechanically wounded
White lines, cell borders
Color bar, approximate [Ca²⁺]i

Intracellular Ca2+ Concentration of ROG Cells in Response to FSH and ATP

: Endocrinology Vol. 141, No. 9 3461-3470.
Courtesy of Dr. T. Ji

Fura-2 AM loaded ROG cells
Illuminator: PTI DeltaRAM
Camera: ICCD camera
Software: PTI ImageMaster™

[Ca2+]i Response to ATP

: Am J Physiol Lung Cell Mol Physiol 280: L221-L228, 2001.
Courtesy of Dr. S. Boitano

Fura-2/AM loaded ROG cells
Illuminator: PTI DeltaRAM
Camera: ICCD camera
Software: PTI ImageMaster™

Regenerative Calcium Oscillations

: Biophys J, 79 (5): 2509-2525, 2000.
Courtesy of Dr. I, Pessah

Fura-2/AM loaded differentiated 1B5 myotubes
Illuminator: PTI DeltaRAM
Camera: ICCD 300 camera
Software: PTI ImageMaster™
Images: (A) Cells stimulated with 3 mM caffeine. After 2 s, a calcium wave begins from a discrete region and spreads across the cell. After ~2 s more, the calcium wave occurs again. (C)The corresponding change in the Fura-2 340/380 ratio (B) Ratio images from the same cell in A stimulated with 40 mM caffeine. Calcium increases globally throughout the cell, and no calcium waves or oscillations are observed. (D) The corresponding change in the Fura-2 340/380 ratio

Simultaneous Measurement of Phagocytosis and [Ca2+]i

: J Cell Sci 2003;116:2857-2865.
Courtesy of Dr. Dewitt, S. et al.

Illuminator: PTI DeltaRAM
Camera: PTI ICCD100
Fura-2 labeled human neutrophils were presented with a DCDHF-labelled C3bi-opsonised particle for phagocytosis
Images: Phase contract (top) and corresponding fura2 signal (middle). 90 s: micropipette presenting the particle to the cell; 102 s:adhesion of the particle to the cell without Ca²⁺signaling; 123 s: formation of the phagocytic cup; 141s: closure of the phagosome 180 s: completion of the event and the return of cytosolic free Ca²⁺ to baseline

Correlation of Oxidative Activation with Ca2+ and Phagocytosis

: J Cell Sci 2003;116:2857-2865.
Courtesy of Dr. Dewitt, S. et al.

Illuminator: PTI DeltaRAM
Camera: PTI ICCD100
Images: Top: Phase contract to show the phagocytic event
Middle row: corresponding fura2 signal to show cytosolic free Ca²⁺ changes.
Bottom row: DCDHF fluorescent intensity of the internalized zymosan particle to assess oxidative activity The graph at the bottom shows the complete time course for cytosolic free Ca²⁺ change (black) and DCDHF intensity (SI) with the point of phagosomal closure marked by the arrow
Conclusion: the onset of oxidative activity correlates with the second phase of the Ca²⁺ signal.

Local Oxidase Activation and Ca2+ Signal Reported by Fura2-dextran Conjugate

: J Cell Sci 2003;116:2857-2865.
Courtesy of Dr. Dewitt, S. et al.

Illuminator: PTI DeltaRAM
Camera: PTI ICCD100
The Fura-2 dextran conjugate micro-injected neutrophils was challenged with an opsonised particle
Images: Phase contract (top) and corresponding Fura-2 dextran signal (bottom) show the phagocytic cup (270 seconds), phagosome closure (340 seconds) and completion of the Ca²⁺signal (380 seconds)
The graph on the right shows the complete Ca²⁺ data, with the point of phagosome closure marked by the downward arrow.

Store-Operated Sr2+ Entry

: J. Biol. Chem., 278 (43):42427-42434,2003.
Courtesy of Dr. J. Deans

Fura-2/AM loaded CHO cells transfected with CD20 or vector
System: PTI DeltaRAM based-ImageMaster™ system
Images: (A) Treatment of the cells
(B) Images at a, b, and c time points:
Before store depletion, no difference in base-line fluorescence, and no Sr²⁺ entry
ATP depleted Ca²⁺ stores and sharply increased [Ca²⁺]c in both cell lines
Subsequent perfusion of Sr²⁺ induced a large increase in the CD20-transfected but not the control cells

Use of DCDHF as an Oxidative Indicator During Phagocytosisy

: J Cell Sci 2003;116:2857-2865.
Courtesy of Dr. Dewitt, S. et al.

Illuminator: PTI DeltaRAM
Camera: PTI ICCD100
Images: (a) DCDHF-conjugated zymosan particles before (left) and after (right) addition of H2O2
The traces below show the time courses for the increase in fluorescence with the arrow indicating the addition of H2O2
(b) Fluorescence intensity of internalized (arrowed) and adherent (asterisk) DCDHF-conjugated zymosan particles
The DCDHF intensity image and the phase contrast image have been superimposed for clarity.

Ca2+ Imaging of Rat Medium Spiny Neurons

: Neuron, 39 (7): 227-239, 2003.
Courtesy of Dr. Ilya Bezprozvanny

GFP expression S2 cells loaded with Fura-2
Illuminator: PTI DeltaRAM
Camera: PTI IC-300
Software: PTI ImageMaster™ Pro

Dose-dependence of Trehalose Response in S2-Gr5a Cells

: PNAS 100 (suppl. 2):14526-14530, 2003.
Courtesy of Dr. Carlson, J

GFP expression S2 cells loaded with Fura-2
Illuminator: PTI DeltaRAM
Camera: PTI IC-200
Software: PTI ImageMaster™
Images: Upper: Divided panels of S2-Gr5a cells (Left and Center) or negative controls, transfected with GFP vector alone (Right), before and after application of either trehalose (Left and Right) or maltose (Center)
Lower: Images of fields of S2-Gr5a cells taken on application of different concentrations of trehalose.

Time Course of Trehalose Response in S2-Gr5a Cells

: PNAS 100 (suppl. 2):14526-14530, 2003.
Courtesy of Dr. Carlson, J

GFP expression S2 cells loaded with Fura-2
Illuminator: PTI DeltaRAM
Camera: PTI IC-200
Software: PTI ImageMaster™
Images: Upper: A series of images of a single fura 2-loaded S2-Gr5a cell, taken at 5 s intervals
Lower: A quantitative representation of the response of the same cell. Bar indicates stimulus period.

Cells Expression Redo-GFP

Redo-GFP expressing cells
PTI DeltaRAM illuminator
Roper Sensy camera
PTI ImageMaster™ 3.0 software

Visualization of Mitochondria by mitoGFP

: EMBO, 18(1): 96-108,
Courtesy of Dr. Hajnoczky

mitoGFP transfected intact (A) and permeabilized (B-D) mast cell
Cells were also loaded with MitoTracker Red (C) or rhod2/AM (D)
PTI DeltaRAM illuminator.
Photometrics PXL CCCD camera
The green images (left panels) show the distribution of mitoGFP, the red images (middle panels) show the distribution of MitoTracker Red (C) or compartmentalized rhod2 (D). These images are overlaid in the right panels to show the coincidence of the labeled organelles (overlay).

IP3-induced Intracellular [Ca2+]c and Mitochondrial [Ca2+]m Responses

: EMBO, 18(1): 96-108,
Courtesy of Dr. Hajnoczky

Fura2FF-loaded permeabilized cell
PTI DeltaRAM illuminator
Photometrics PXL CCCD camera
Left: the overlaid images show the distribution of the membrane-bound CaGreen-C18 (image i, purple) and the mitochondrially compartmentalized Fura2FF (image i, green), and the changes in the Fura2FF fluorescence (images ii-v, 380 nm green/340 nm red) upon addition of 100 nM IP3 (ii versus iii), 12.5 M IP3 (iii versus iv) and ionomycin (iv versus v)
Right: time courses of the global [Ca²⁺]pm response (vi) and the average [Ca²⁺]m response (vii, thick line), and the [Ca²⁺]m responses of the marked (1–6 on image i) individual mitochondria (vii, thin lines).

Simultaneous measurement of Ca2+ and myocyte cell length

: Dual emission channel OBB photometer for Indo-1 Ca++ measurements

The above data traces are from the simultaneous collection of fluorescence and cell length from a cardiac myocyte. Myocyte was loaded with dual emission ratiometric fluorescence probe Indo-1. The OBB/PTI DeltaRAM illuminator was used for excitation illumination at 365 nm and the emitted fluorescence was detected with a dual emission PMT photometer. Above data was collected with FeliX32 photometry software from Photon Technology International (PTI). The blue trace shows the calcium ratio increase and decrease with the cell contraction. The contraction data (Green trace) was detected with a video edge detection electronics module and is correlated with the fluorescence data since both signals were collect simultaneously

Simultaneous Ca++ Fluorescence and Patch Clamp Electrophysiology Research

: Image courtesy of, Prof. Dr. György Panyi, Department of Biophysics and Cell Biology, University of Debrecen

Shown is a picture of a PTI/OBB dual emission photometer attached to the C-mount of an inverted fluorescence microscope. The microscope is also equipped with electrophysiology recording devices and perfusion apparatus. The entire setup is inside a Faraday cage for electrical isolation.

OBB’s photometers are ideal for quantitative live cell measurements of intracellular ion and molecules such as Ca⁺⁺, Na⁺⁺, pH, GFP, FRET and FRAP experiments. By allowing for simultaneous quantitative fluorescence detection with patch clamp recordings, these photometers have been widely used by electrophysiologists around the world. The OBB PMT photometer is a passive detection system that provides an output voltage signal proportional to the ion/fluorophore of interest, and this signal is directly fed into the electrophysiology A/D converter and collected with the customer’s existing software. The OBB photometer can collect a fluorescence signal at up to 20 KHz for high speed transient recordings.

Applications Notes

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