The new LABION Laboratories accommodate several instruments and equipment required for responding to several diagnostic and research issues by using different approaches.
Indeed, the laboratories are equipped with both traditional methods used in chemistry, biochemistry, molecular biology and histology, together with more innovative biophotonic instruments like the Raman microscopes.
Raman spectroscopy is a non destructive vibrational spectroscopy able to provide the chemical composition of the analyzed sample by a simple irradiation with laser light. Differently from IR, Raman spectroscopy is not affected by the presence of water and therefore is particularly attractive for biomedical studies.
Aramis (Horiba Jobin-Yvon) is a state of the art confocal micro-Raman spectrometer fully controlled through Labspec 6 software with advanced imaging functionalities. The system is equipped with three different laser lines operating at at 532 nm (500mW), 633 nm (50 mW) and 785 nm (500 mW) and has three interchangeable gratings with 600 l/mm 1200 l/mm, 1800 l/mm. The maximum spectral resolution is of about 1 cm-1 wavenumber using the 1800 l/mm grating.
Surface Plasmon Resonance imaging
LABION is the first world wide user of XelPlex (Horiba Jobin-Yvon), a new high-performance and full-automated instrument based on Surface Plasmon Resonance imaging (SPRi). SPRi is based on the direct and label-free detection of the interaction between two molecules (e.g. proteins, smallpeptides, antibodies, drugs, DNA, vesicles, bacteria). The first molecule (“ligand”) is normally immobilized on the gold surface of the SPRi-chip and can then capture the molecule of interest (“analyte”) contained in the sample (e.g. blood, serum, urine). Taking advantage of small changes in the refractive index of the gold surface illuminated by a laser, the SPRi can directly measure the interaction between the two molecules. EzPlex allows the concomitant and fast multiplex detection of hundreds of different analytes in multiple samples and biologic fluids to monitor simultaneously and efficiently several biomarkers related to the disease status, the efficacy of therapeutic treatments and the effect of rehabilitation.
Mass spectroscopy (MALDI-TOF)
The MADLI-TOF (Bruker Daltonics Microflex LT) is a bench-top mass spectrometer used for the analysis of proteins, other small molecules (lipids, carbohydrates) and microorganism (bacteria). Thanks to this instrument is possible to perform fast and reliable analysis for biomarker discovery and for diagnostic purposes. In addition to the high mass accuracy and sensitivity, one of the most relevant advantages of this instrument is the easy, fast and reproducible sample preparation.
PCR and related molecular biology techniques
The PCR is the universal technology used for the amplification of a genetic sequence (DNA) derived from both real clinical samples or from synthetic processes. This method can be used for several research and clinical purposes, in particular, in our laboratory PCR is used for the production of oligonucleotides modified in order to be coupled to nanostructured components and as golden standard technique for comparing and validating the performances of innovative biophotonics/nanotechnology-based methods developed in our laboratory.
- DNA Extraction
- RNA Extraction
- cDNA production
UV/VIS Spectrophotometer (Nanodrop)
The NanoDrop 2000 is a wide spectral range (190-840 nm), easy to use UV-Vis Spectrophotometer for a big variety of applications including analysis of peptides, DNA, RNA, protein, nanoparticle samples, colorimetric assays, dyes. Works with normal and micro plastic and quartz cuvettes or using the nanodrop function requires only 0.5 – 2 µL of the sample. The user friendly software includes pre-configured methods for DNA, Protein A280, Microarray, Protein and Labels, Pierce 660, Bradford, BCA, and Lowry and has various data export capabilities.
High-performance liquid chromatography (HPLC) is a typical multipurpose analytical method for the separation and identification of several class of molecules. This instrument can be used for both the identification of biomarkers in clinical samples (e.g form blood, serum, urine, tissue homogenate) and for the identification of compounds derived from chemical reactions or cells cultured in-vitro.
Scanning electron microscope (SEM)
The Hitachi TM3000 table top SEM enables taking good resolution images without special sample preparation in a very short time. The low vacuum mode allows imaging of hydrated or non-conducting samples and two selectable beam energies (acceleration voltage) help imaging of soft or hard matter. A high-quality multi-segment electron detector provides contrast-rich images with selectable material/compositional or topographical emphasis. Thanks to the simplicity of the software, users can fast become independent operators after short training. The large stage allows introduction of vide variety of samples related to life science, food, cosmetics, healthcare, pharmaceutical, textiles or materials science.
Franz-type diffusion cell
The Franz Cell is a simple, reproducible test for in-vitro skin permeation studies. Franz-type system is relatively simple in the design (see the scheme below), the cell comprises two parts: the donor chamber that contains the substance (e.g. drug , nanoparticles) to be tested and the receptor chamber containing the medium with the substance diffused and the sampling port. The two parts are separated by a membrane, human skin in our experiments. The receptor temperature is usually maintain to 32 degrees C°, to approximate normal skin conditions, thanks to a heater circulator.
The samples are collected for several hours and analysed using HPLC or similar technique.
LABION is equipped with a complete system of 6 vertical Franz-type diffusion cells (Permegear), stirrer station and thermostated water machine (see the figures below).
Histology instruments and services
Histological analysis is one of the most widespread techniques in life science research, useful both for diagnostic purposes and basic biomedical studies. Chemical fixation and paraffin embedding of tissue samples allow long lasting preservation of the physiological morphology and the molecular composition of the biopsy. Taking advantage of a manual rotary microtome, it is possible to obtain serial thin paraffin sections (2-10 µm) suitable for subsequent histological staining and immunolocalisation techniques. Routine staining procedures guarantee the observation of the overall tissue cytoarchitecture, whereas structural protein expression might be highlighted by means of multistep immunological reactions.
In our laboratory, frozen samples can be cryosectioned thanks to a cryostat designed to prevent abrupt temperature variation of the sample while sectioning. High resolution, bright field images are obtained with an upright light microscope equipped with digital camera.
The Leica DM2500 upright microscope is designed for the observation of histological sections in bright field, differential interference contrast, polarization contrast, and high-performance fluorescence. Chromatically-corrected optical paths ensure high contrast images from 25x to 630x magnification. For epi-fluorescence microscopy, thanks to three integrated filter blocks, it is possible to select filters and dichroic mirrors according to the fluorophore used, thus obtaining the optimal fluorescence signal with minimal background light. The camera associated software with a multi-channel imaging capability allows instant high resolution image overlay, morphometric analysis, and image processing.
Biological safety cabinet
The biological safety cabinet ensures the aseptic or particulate free conditions needed for tissue and cell culture experiments. It uses filters and directional horizontal laminar air flow to provide a contaminant-free work area. The horizontal flow hood belongs to the Class II biological safety cabinets which protect the specimen, the user, and the environment from contamination and it is suitable for applications requiring sterile conditions without the need to work with volatile chemicals.
Laboratory chemical hood
Refrigerated Centrifuge SIGMA 3-16PK