1. FT-IR microspectroscopy: a powerful tool for spatially resolved studies on supports for solid phase organic synthesis
Lisa Vaccari

2. Outline Source for Imaging and Spectroscopic Studies in the Infrared
Beamline layout
MidIR experimental station
FT-IR Microspectroscopy
Solid phase organic synthesis
Introduction
Reaction Kinetic
Diffusion Process
Acknowledgements

3. Layout of SISSI
Source for Imaging and Spectroscopic Studies in the Infrared
Radiation is collected over a solid angle of
65 mrad (H) x
25 mrad (V)
M1 Plane mirror
M2 Ellipsoidal mirror
M3 Plane mirror
M4 Ellipsoidal mirror
a = 3.5 m d = 1.5 m
b = 1.0 m e = 1.0 m
c= 11.5 m f= 2.5 m d c a b e f
M1 Plane mirror
M2 Ellipsoidal mirror
M3 Plane mirror
M4 Ellipsoidal mirror

4. Experimental stations
Switching
Mirror (M5)‏
2nd branch
1st Branch (CNR-INFM)‏
Solid State Physics
High Pressures
Time-resolved spectroscopy
2nd Branch (Elettra)‏
Biophysics/Biochemistry
Spectroscopy and Imaging
VERTEX
Hyperion
3000
IFS66/v
Hyperion
2000
1st branch

5. MidIR Microspetroscopy
Visible
Microwaves
Mercury-Cadmium-Telluride Detector
Active area of 250X250 mm2
Operation range: cm-1
Focal Plane Array detector
64X64 pixels 2.5X2.5 mm2 active area
Operative range: cm-1

6. Chemical Imaging
Generate Image Contrast by Using Vibrational Spectral Properties
Chemical Sample Mapping
Chemical Sample Imaging
Vibrational spectra of a sample point by point irradiating small sample areas
Vibrational spectra of many sample points Irradiating the full field of view
Single point MCT detector
64X64 pixels of FPA detector

7. Diffraction Limited d = 0.61 l / NA
Lateral Resolution
Diffraction Limited d = 0.61 l / NA
Objective NA
Wavelength d
15X
0.4
10 mm (1000cm-1)
15 mm
2.5 mm (4000cm-1)
 4 mm
36 X
0.5
12 mm
 3 mm
FPA Detector
Objective
Pixel resolution
15X
 2.6
36X
 1.1

8. Acquisition Time vs Sensitivity
MCT Detector
Scan Velocity 20 KHz
Number of scans 32
Spectral resolution 4 cm-1
2.36 spectra per second
SNR  10-5 au
Easily usable with SR
FPA Detector
Scan Velocity 6 KHz
Number of scans 32
Spectral resolution 4 cm-1
3048 spectra per second
SNR  10-3 au
Special applications with SR

9. MIR Performance of SISSI
FPA detector
32 scan
Res:8 cm-1
MCT detector
128 scan
Res:4 cm-1
Development of Globar-FPA/Synchrotron-MCT combined approach
Fast acquisition of sample images with FPA detector to check sample quality and to identify regions of interest
Higher quality map collection exploiting the brightness advantage of SR and major sensitivity of MCT detector

10. Solid Phase Synthesis
Large compound libraries of peptides, oligonucleotides and small molecules (drugs)
Inert resin support
I block
II block
Product
Distribution of reaction products into the bead can gives information on pore wettability and accessibility, efficiency of the reactant diffusion process, load capacity of the bead and reaction kinetics
BEAD PERFORMACES

11. Optical Transparent Polymeric Resins
Annie Y. Bosma, Rein V. Ulijn, Gail McConnell, John Girkin, Peter J. Halling and Sabine L. Flitsch
Using two photon microscopy to quantify enzymatic reaction rates on polymer beads Chem. Commun., 2003, 
Non-Optical Transparent Polymeric Resins
ATR powder
Flatten Single bead microscopy
ATR Microscopy

12. Average pore diameter (nm)
Synbeads
Rigid methacrylic polymeric beads
Non-swelling and rigid support – High mechanical stability
Versatile - Controlled porosity and different chemical functionalities
Recyclable
Synbeads type
Average pore diameter (nm)
30 ÷ 40
80 ÷ 100
200 ÷ 250
Amino-Methacrylate
A110
A210
A310
Carboxyl-Methacrylate
X110
X210
X310
Hydroxymethyl-Methacrylate
H110
H210
H310
Chloromethyl-Methacrylate
C110
C210
C310
In collaboration with Pharmaceutical Science Department of Trieste University (Prof. L. Gardossi, A. Basso, S. Cantone, L. Sinigoi) and Resindion Mitsubishi Chem. Corp. (Milano)-

13. Test reaction - kinetic - 1 -
Amino-Methacrylate beads – A310 – average pore diameter of nm
Reaction time:5,10,20,30,40,60 min; 1eq polymer- 3 eq nitropropionic acid; Bead loading: 0.85 mmol/ gr dry
Evanescent field propagation
Sample Ge
Detector
Source
Ge (n1=4),  = 45˚, organic medium n2 = 1.5
dp (1550cm-1) = 428 nm

14. Test reaction - kinetic - 2 -

15. Test Reaction – Diffusion - 1 -
Infrared Microscopy is a label free assay
5μm thin bead sections
Average diameter μm
FPA Images
64 scans, 4cm-1

16. Test Reaction – Diffusion - 2 -
FPA Images, 64 scans, 4cm-1
Reaction time: 10 min
Reaction time: 30 min
Reaction time: 60 min

17. Test Reaction – Diffusion - 3 -

18. SR-FTIR Microspectroscopy. 5μm spatial resolution, 256 scans, 4cm-1
Test Reaction – Diffusion - 4 -
SR-FTIR Microspectroscopy. 5μm spatial resolution, 256 scans, 4cm-1
10 min
20 min
30 min

19. Conclusion and future developments
We propose a new approach for spatially resolved studies of chemical distribution based on the combination of two FTIR microscopic techniques: Conventional Source-FPA/SR-MCT detector
The high spatial resolution and fast acquisition time of FPA detector are exploited for a rapid screening of the samples to identify the best ones to be measured
The major sensitivity of MCT detector and high brightness of SR source are exploited to highlight spectral features otherwise not easily detectable
The proposed approach is sensitive and fast enough to be employed for a systematic study of reaction kinetics and diffusion mechanism for solid phase chemistry and to be extended to others scientific problems
0 min
6 min

20. Acknowledgements Thanks for your attention
Trieste University – Pharmaceutical Science Department
Prof. Lucia Gradossi, Alessandra Basso, Sara Cantone and Loris Sinigoi
SISSI group
M. Kiskinova, D. Eichert, F.Morgera G. Birarda and D. Bedolla
S. Lupi, A. Perucchi, R. Sopracase,
Thanks for your attention