Dr. ZherdevaV., Dr. Loginova Y., Dr. Kazachkina N., Dr. Savitsky A. Bach Biochemistry Institute of Russian Academy of Science Moscow 2013

Semiconducter core: Cd/Se, Cd/Te, и Ga/N shell: Zn/S, Cd/Se biofunctional coating: polymer, protein or lipid  Excitation in broad range of spectrum  emission depends on size  Lifetime of ns  high extinction coefficient  high quantum yeild  uniqe photostability - Semicondacter nanocrystal (1-10 nm) with unique photophysical properties :

APPLICATION OF QDS Michalet X, Pinaud FF, Bentolila LA, Tsay JM, Doose S, Li JJ, Sundaresan G, Wu AM, Gambhir SS, Weiss S: Quantum Dots for Live Cells, in Vivo Imaging, and Diagnositics. Science (5709):

QDs is a perspective tools for medical usage… TOXIC or NON TOXIC? Distribution Localization and accumulation in vivo. Distribution Localization and accumulation in vivo. per osIntravenously (i.v.)

TECHNOLOGY Type of administration I.V. Per os I.V. Per os Localization study In vivo imaging Local fluorescence spectroscopy Confocal fluorescent microscopy Pathomorphogy Morphology Clinical biochemistry Toxical effects

QDs MPA λ em 611nm and 630 nm d ~ 8 – 11нм QY % QDs PolyT λ em 626 нм d ~ 15 – 16 нм QY 10-30% QDs PolyT-APS λ em 678 нм d ~ 36 нм QY 5-20% QY

FLUORESCENT EQUPMENT iBox UVP Laser spectrometer with optical zond

Monitoring of fluorescence in vivoin digestive tract of mice Excitation nm, emission filter nm exposition25с. per os

QDs Stomach after 2 h Intenstines after 2 h Intenstines after 4 h Intenstines after 6 h MPA+--- PolyT++±- PolyT-APS++++ No fluorescence (-), weak (±), high (+). Local fluorescense spectroscopy Fluorescence of excrements after 24 h of QDs adminestering per os QDs MPA Black line- excrements of control mice Red line- excrements of after QDs administering QDs PolyT-APS QDs PolyT

I.V. QDs MPA LFS

QDs Poly T LFS I.V.

i.V. QDs PolyT-APS LFS

Organs Heart Lungs Liver Spleen Kidney Brain Limp nodes М uscels Skin Intestines Fluorescence after 40 min +* * + Fluorescence after 22 days Confocal microscopy NB : weak +, medium ++, high +++, no -. (*) in blood vessel only а – reflecte lights, б – fluorescence, в – peudo color imaging, г – fluorescence spectra Lungs Liver а Lungs

а – local haemorragia; б – stenosis of arteria.  Blood Urea creatinine,  Morphology Toxicity of QDs MPA No change alkaline phosphatase alanine - and aspartate transaminases Hihg spleen weight (twice )  Pathomorphology а б

I.V. QDs MPA toxicity: also depends on initial solution

1.Distribution and accumalation depend on the modification of coating 2.Qds are dissolving in digestive tract: PolyT-APS are more stable comparing to QDs МPА and PolyT after per os administering 3.The main targeted organs for QDsМPА are the lungs, for PolyТ are the atria, for PolyT-APS are liver after I.V. Excretion of in the urine and faeces was not detected. 4.QDs of small size МPA stay in organism for a long period.They could lead to long-term pathomorphological changes in lungs on the 22-th day after administration. 5. The the severity of the toxic effect of QDs MPA depends on the solvent CONCLUSION

Локальная флуоресцентная спектроскопия (ЛФС)