LABORATORY OF SPECIAL MICROSCOPIC TECHNIQUES (L-7) |
Scope of accreditation of research laboratory No AB 120
issued by Polish Centre for Accreditation Issue No. 17 of 12 June 2019.
Head of the laboratory | The experts |
Roman Major PhD, Eng. | Przemysław Kurtyka, MSc. Eng. |
Address:
Institute of Metallurgy and Materials Science Polish Academy of Sciences
ul. Reymont 25, 30-059 Krakow
phone: (48) 12 295 28 98, fax: (48) 12 295 28 04
e-mail: Adres poczty elektronicznej jest chroniony przed robotami spamującymi. W przeglądarce musi być włączona obsługa JavaScript, żeby go zobaczyć. , website: http://www.imim.pl
Apparatus:
Confocal Laser Skaning Microscopy LSM Exciter 5 with the incubation chamber
• Confokal Modul LSM 5 Exciter, two canals, RGB
• Laser HeNe 633nm 5mW
• Laser HeNe 543nm 1mW
• Laser argon 458/488/514nm, 25mW
• Diode laser V 405nm
• Main Beam Splitter turret PASCAL
• Software ZEN 2008 LSM 5 EXCITER
• Light division system (405, 458, 488, 514, 543 nm)
• Filter BP 505-530
• Filter BP 505-600
• Filter BP 530-600
• Filter BP 560-615
• Filter LP 420
• Filter BP 420-480
• System ECU LSM 5 EXCITER
• Modul DIC I/0,9 z polaryzatorem
• Transmited light detectorT-PMT LSM 710
• Heating stage
• Incubation system
Figure 1 Confocal Laser Scanning Microscopy (working in IMMS PAS)
Research possibilities:
It is a fully automated system with full control of the microscope setup, the operating parameters of lasers and stepless confocal aperture. Its key features include the ability to register up to 6 independent channels and their overlap, MULTITRACKING option that allows registration-free phenomenon of cross-talk with overlapping issues, ease of use thanks to the muli- excitation function REUSE software and recording of fast processes. The control software ZEN 2008 (Deconvolution ZEN 2008, Package Physiology ZEN 2008, Package Topography ),
and calculation software Axio Vision 4.8 (Pakiet oprogramowania AxioVision 4 Module AutoMeasure
AxioVision 4 Module AutoMeasure
Automatic measurements. Executive generator:
- Basic functions of image processing - Total or local segmentation boundaries of objects using thresholding histogram
- Automatic segmentation of objects and interactive processing of the measurement masks
- Geometric and densitometric measurements of individual objects
- Selecting objects measured, presentation of results in the image plane
- Saving images in the format *. CSV files compatible for Excel
- Performing measurement programs
- Clustering processes for any number of images
- Activating and deactivating as well as changes in the parameters measured during the analysis
The software package AxioVision 4 Module AutoMeasure Plus.
Among the examples of the application of the confocal microscopy is the 3D analysis of cells and tissues, multi-channel recording with quantitative measurements phenomenon co location, monitoring of physiological processes
Figures 2a-c show the successive steps of a confluent forming:
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Figure 2a After one hour | Figure 2b After 24 hours |
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Figure 2b After 72 hours | |
Scanning Acoustic Microscopy SAM
The acoustic microscope works in the pulse reflection method. The most important component in the scanning acoustic microscopy is a high frequency piezoelectric sound transducer. This object transmits and receives sound pulses of high penetration rate. It is a sapphire cylinder with a ZnO film (1). A transducer generates a ultrasound pulse (piezoceramic layer converts electromagnetic vibrations into sound wave) which propagates along the delay rod. The transducer is immersed within a coupling medium (water). The immersion system cavity is therefore the acoustic spherical lens. A lens focuses beam within the sample. The acoustic objective receives the sound reflections from the sample. The information follows to the return to transducer. There transforms them into electromagnetic pulses detected by an oscilloscope and on the monitor display them as a pixel. The acoustic objective scans the sample line by line. Figure 3 shows a scheme of an acoustic microscope system.
Figure 3. Scheme of the Scanning Acoustic Microscope
The result of the SAM is determined by aperture and acoustic wavelength, which is depends on the material but this microscope can detected damage to a minimum size of 0,3 µm. Depending on the critical defect size and defect depth the working frequency and transducer design have to be chosen. The important information is that the SAM works in real time mode. The scanning acoustic microscope is equipped with a 4 heads (15MHz, 75MHz, 110MHz, 180MHz).
The Scanning Acoustic Microscope works on several modes (Figure 4). A 2D scan gives information about x-y plane and 3D scan image provides information about x-y plane and the time of flight the acoustic beam. The most popular are scan A, scan B, scan C and scan X. Scan A mode is used to characterize a single point of interest in a specimen. The B-scan mode generates a top-down or X-Z axis image. The C-scan is a compilation of A-scan but not in one point but along X-Y axis, they a display of the image of reflected echoes at the focused plane of sample. Scan X is used to view image at a depth of the sample.
Figure 4. Scan mode in Scanning Acoustic Microscope
Example
Having described the principles of the scanning acoustic microscope is now presented same results. The scanning acoustic microscopy is being used to analyse: bonded and soldered structures, castings, interface and semiconductor components, material stress and crack propagation, interface evaluations of thin coatings, biological, geological and ceramics structures, detection of delaminations in packages, 3-dimensional imaging of welding, determining volume defects .
One of the applications of the scanning acoustic microscopy was in the study of bio-ceramics mater. In Figure 5 is presented an acoustic image of hydroxyapatite sintered at various temperatures. This material is characterized by inhomogeneous and different pores sizes.
Figure 5. Visualization of micropores or inhomogeneous material (hydroxyapatite) using the SAM
The SAM is a good techniques to detect small defects in the surface and under the surface. Using this technique it is possible to find cracks smaller than 1 µm. Surface near cracks, delaminations, void or inclusions due to their different elastic properties from the surrounding materials. In figure 6 is presented an acoustic identification of the defects within the massive forging ingot. The SAM identification of defects at difference depth from the surface. In the figure 6 are visible defects (holes), contains a carbon stalagmite (yellow arrow indicates the carbide localized inside the void) and some porosity. In figure 7 is presented a deeper layer in this material (forging ingot).
Figure 6. Visualization of carbides localized inside the void and common defects (porosity) in ingot
Figure 7. Image to the sample forging ingot applying the method "layer by layer" from the surface registered by the SAM
Contact for the Client:
Head of the Lab L-7:
Roman Major, PhD, D.Sc.
phone: +48 12 295 28 41/05
e-mail:
Adres poczty elektronicznej jest chroniony przed robotami spamującymi. W przeglądarce musi być włączona obsługa JavaScript, żeby go zobaczyć.
