2011.11.25.. TÁMOP – 4.1.2-08/2/A/KMR-2009-0006 1 Development of Complex Curricula for Molecular Bionics and Infobionics Programs within a consortial* framework**
Consortium leader
PETER PAZMANY CATHOLIC UNIVERSITY
Consortium members
SEMMELWEIS UNIVERSITY, DIALOG CAMPUS PUBLISHER
The Project has been realised with the support of the European Union and has been co-financed by the European Social Fund ***
**Molekuláris bionika és Infobionika Szakok tananyagának komplex fejlesztése konzorciumi keretben
***A projekt az Európai Unió támogatásával, az Európai Szociális Alap társfinanszírozásával valósul meg.
Peter Pazmany Catholic University Faculty of Information Technology
Neurobiológia alapjai - Módszerek
BASICS OF NEUROBIOLOGY - Methods
www.itk.ppke.hu
By Imre Kalló
2011.11.25. TÁMOP – 4.1.2-08/2/A/KMR-2009-0006 3
METHODS IN NEUROBIOLOGY IX.
Dissection, virtual dissection, imaging techniques
Imre Kalló
Pázmány Péter Catholic University, Faculty of Information Technology
I. Histology techniques: light microscopic studies II. Applications using fluorescent dyes
III. Histology techniques: electron microscopic studies IV. Techniques to map neuronal connections
V. Molecular biological techniques VI. Living experimental models VII. Electrophysiological approaches VIII. Behavioral studies
IX. Dissection, virtual dissection, imaging techniques
Basics of neurobiology: Methods in Neurobiology IX.
www.itk.ppke.hu
DISSECTION, VIRTUAL DISSECTION, IMAGING TECHNIQUES
„Mortui docent vivos”
Classical way of dissection - in situ demonstration of the anatomical and pathological details in their 3D appearance – for specialist
New preservation technique – plastination makes the whole body or body parts possible to preserve for long time and to demonstrate them with safety – also for public
New approach – high resolution maps of the human body accessable through user friendly softwares
New elements – human atlas of 2-8mm-thin slices cut from epoxy embedded bodies in the major planes of space - to support the new imaging techniques
TÁMOP – 4.1.2-08/2/A/KMR-2009-0006 5 2011.11.25.
DISSECTION, VIRTUAL DISSECTION, IMAGING TECHNIQUES
„Viva docent vivos”
Classical way of imaging – 2D projection -, reflection -, emission images of the body or body parts from a single direction
„New” imaging techniques – based on tomography - gathering projection data from multiple directions
Modern tomography – combined with high speed
computation to process, analyse, combine, store images and make 3D reconstructions of the body or body parts.
X-ray
CT MR
PET SPECT
New approach in neuro research – new tools to investigate population of neurons activated by defined cognitive tasks.
fMRI
Basics of neurobiology: Methods in Neurobiology I.
www.itk.ppke.hu
IMAGING TECHNIQUES
Photography Radiation Electro-magnetism
Microscopy X-ray MRI
Optical imaging SPECT (MEEG)
PET (BEAM)
Autoradiography
TÁMOP – 4.1.2-08/2/A/KMR-2009-0006 7 2011.11.25.
IMAGE SLICE – PIXEL - VOXEL
pixel: picture element (2D) voxel: volume element (3D) dimensions:
x, y, z and (value)
Basics of neurobiology: Methods in Neurobiology IX.
www.itk.ppke.hu
IMAGING TECHNIQUES: PET
Isotope-production:
11C, 13N, 15O, 18F
Cyclotron Radiochemistry
PET scanner
Image processing and analysis
isotope
radioligand
Quality control
TÁMOP – 4.1.2-08/2/A/KMR-2009-0006 9 2011.11.25.
IMAGING TECHNIQUES: PET
Parameters (biochemical – physiological) measured
Blood flow, blood volume Metabolism
oxygen, glucose, aminoacids, fatty acids, fluor, ...
Receptor systems:
dopaminergic, cholinergic, serotoninic, benzodiazepin-GABA, opioid, ...
Pharmacodynamics and pharmacokinetics Protein synthesis
Molecular diffusion
Tissue pH
Basics of neurobiology: Methods in Neurobiology IX.
www.itk.ppke.hu
IMAGING TECHNIQUES: PET
Parameters (biochemical – physiological) measured
CMRglu CBF GABA
Amethionine
TÁMOP – 4.1.2-08/2/A/KMR-2009-0006 11 2011.11.25.
IMAGING TECHNIQUES: MRI
B0 M0
B1 Proton spins of the subject in the big magnetic field
(B0) are aligned parallel or antiparallel with the direction of the field.
Radio waves are transmitted into the subject [for about 3 ms] – perpendicular to B0, the magnetic component (B1) of this electromagnetic wave temporarily knocks the protons out of alignment
Radio wave transmitter is turned off, while it receives radio waves re-transmitted by the protons ‘relaxing’
back to their undisturbed ‘equilibrium’ position. Re- transmitted radio waves are manipulated with magnetic fields during this readout interval [10-100 ms]
Storing of measured radio wave data vs. time – repetitions of transmission- reception
Processing raw data to reconstruct image
B0
B0
Basics of neurobiology: Methods in Neurobiology I.
www.itk.ppke.hu
IMAGING TECHNIQUES: MRI Comparison of T1 and T2 modes
TÁMOP – 4.1.2-08/2/A/KMR-2009-0006 13 2011.11.25.
IMAGING TECHNIQUES: fMRI
Physiological parameter measured:
Increased neuronal activity requires more glucose and oxygen , which is rapidly delivered through the blood stream resulting in local changes in cerebral blood volume (CBV) and cerebral blood flow. These in turn generate local changes in the relative concentration of oxygenated (HBO2) and deoxygenated hemoglobins (HBr), which serves as the Blood
Oxygen Level Dependent signal (BOLD), a marker for MRI (hemoglobin is diamagnetic when oxygenated, but paramagnetic when deoxygenated.
Basal state Arteriole Activated state
Venule Capillary
bed
Capillary bed
Oxyhemoglobin Deoxyhemoglobin
normal flow basal level (HBr) basal CBV
normal MRI signal
increased flow
decreased level (HBr) increased CBV increased MRI signal
Basics of neurobiology: Methods in Neurobiology IX.
www.itk.ppke.hu
IMAGING TECHNIQUES: fMRI
Shulman R G et al. PNAS 2009;106:11096-11101
Forepaw stimulation in deep anesthetized states results in more localized activation in the
contralateral primary somatosensory cortex.
