Wielofunkcyjny mikroskop 3D Ramana Confotec CARS

 

Wielofunkcyjny mikroskop 3D Ramana Confotec® CARS łączy funkcjonalność:

  1. mikroskopu skanującego CARS
  2. skanującego mikroskopu konfokalnego Ramana/luminescencji
  3. standardowego skaningowego mikroskopu konfokalnego

Model ten umożliwia równoczesny pomiar w następujących trybach:

  1. F-CARS
  2. E-CARS & Raman
  3. Odbitego światła lasera
  4. Przechodzącego światła lasera
  5. Luminescencji laserowej

Zalety techniki CARS

  • high sensitivity: CARS generates more intensive and directed signals in comparison to spontaneous Raman microscopy;
  • anti-Stokes CARS signal has frequency exceeding pumping waves frequencies and is detected in a spectral range free from the stray light of Stokes luminescence;
  • CARS signal is registrated only in a focus where excitation intensity is the highest. It allows imaging with a high spatial resolution using non-confocal pinholes and also performs 3D layer-by-layer scanning with minimal neighboring layers influence on measuring results;
  • spectral resolution of CARS signal is defined only by the width of pumping lasers lines, what simplifies spectral measurements, as detection of CARS signals can be performed without any spectral instrument;
  • CARS signal is proportional to the squared molecule concentration, it allows using CARS (along with the selectivity and noninvasivity of the method) for quantitative measurements of chemical substance concentration in a sample;
  • minimal invasive (nondestructive) CARS method for biological samples. Due to the high sensitivity of CARS method molecules in living cells can be detected without fluorescent markers.

Funkcje

High spatial resolution:

CARS XYZ < 0.7 μм
Raman XY < 300 nm
Z < 700 nm

Wide spectral range:

CARS 985 – 5000 cm-1
Raman 75 – 6000 cm-1

 

High spectral resolution:

CARS 7 – 8 cm-1
Raman 0.25 cm-1

 

Jednoczesne i wielofunkcyjne analizy:

  • szybkie obrazowanie CARS (bez dodawania barwników do próbki)
  • ramanowskie obrazowanie konfokalne
  • obrazowanie konfokalne z dwu lub wielofotonową emisją
  • obrazowanie konfokalne w odbitym świetle lasera
  • obrazowanie z wysokim kontrastem w przechodzącym świetle laserowym
  • wizualizacja profilu próbki przez sygnał drugiej harmonicznej

 

Trzy rodzaje pomiaru CARS:

  • F-CARS
  • Е-CARS
  • P-CARS

 

Five independent speed channels for simultaneous detection up to four 2D and 3D images:

  • F-CARS: CARS signal in forward direction
  • E-CARS & Raman: CARS signal in backward (EPI) direction/ Raman signal
  • Reflected: reflected laser radiation signal
  • Transmitted: transmitted laser radiation signal
  • Luminescent: transmitted laser radiation signal
  • Polarization control over the excitation and detection
  • Mono-block laser system for CARS signal excitation
  • Additional laser 633nm for excitation of a conventional one-photon fluorescence and spontaneous Raman scattering
  • PC-controlled: switching of modes of measurement is performed by automatic switching of the components inside the system

 

Three scanning modes:

  • Laser beam scanning over fixed sample surface with XY scanner
  • Sample transfer by means of XY automated stage relative to the fixed laser beam
  • Multiplexed mode for panorama imaging with a high speed and high spatial resolution: XY scanner + automated stage

Highly-precise calibration over the wavelengths: better than ±0,002nm due to an embedded calibration lamp as a source of reference lines for automated operative calibration of the monochromator-spectrograph.
Block, rigid framing provides a high temporal and thermal stability.

Obraz z mikroskopu 3D Confotec

Multimodal CARS/TPEF image of growing cancer cells HeLa: DNA/RNA/Proteins/Lipids colored

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