Fixed-effects SEM of prostatic stones.

<p>Red arrows show bacterial imprints. a) prostatic carbapatite-amorphous carbonated calcium phosphate (CA-ACCP) stone without any visible imprint; b) spherical prostatic CA stones without any visible imprint; c) prostatic CA-ACCP stone; d) prostatic ACCP stone, e) prostatic CA stone; f) prostatic CA-whitlockite stone. Bar, 4 µm.</p

Mapping and optical image of a birefringent structure.

<p>The poorly soluble foscarnet can be detected and quantified after a mapping of the biopsy thanks to the characteristic peak at 936 cm^-1.</p

Optical image and mapping of BR165 biopsy (scale from blue to red with increasing concentration), and FT-IR spectra of crystals.

<p>a) Optical image of BR165, b) carbapatite map (done at 1030 cm^-1), c) sodium hydrogen urate monohydrate map (done at 3600 cm^-1), d) FT-IR spectra of those compounds.</p

Typical optical image and mapping (scale from blue to red with increasing concentration), and FTIR spectra of crystals generated in Pi 4 or PiCa samples without or in presence of a total concentration of 2 mM of Mg²⁺.

<p>Ia) Optical image, and corresponding Ib) typical FTIR spectra, Ic) Optical image and corresponding Id) typical FTIR spectra of sample PiCa. IIa) Optical image and corresponding IIb) typical FTIR spectra, IIc) Optical image and corresponding IId) typical FTIR spectra of sample PiCa Mg2.</p

Identification of the samples studied through µFTIR experiments.

<p>Samples nomenclature</p><p><b>Ct</b> = samples incubated in 1% FBS DMEM</p><p><b>Pi3</b> = samples incubated in 1% FBS DMEM with Pi set to 3 mM</p><p><b>Pi4</b> = samples incubated in 1% FBS DMEM with Pi set to 4 mM</p><p><b>Pi3 Mg2</b> = incubated in 1% FBS DMEM with Pi set to 3 mM and Mg²⁺ set to 2 mM</p><p><b>Pi4 Mg1.5</b> = incubated in 1% FBS DMEM with Pi set to 4 mM and Mg²⁺ set to 1.5 mM</p><p><b>Pi4 Mg2</b> = incubated in 1% FBS DMEM with Pi set to 4 mM and Mg²⁺ set to 2 mM</p><p><b>Pi4 Mg5</b> = incubated in 1% FBS DMEM with Pi set to 4 mM and Mg²⁺ set to 5 mM</p><p><b>PiCa</b> = samples incubated in 1% FBS DMEM with Pi set to 3 mM and Ca²⁺ set to 2.4 mM</p><p><b>PiCa Mg1.5</b> = samples incubated in 1% FBS DMEM with Pi set to 3 mM, Ca²⁺ set to 2.4 mM and Mg²⁺ set to 1.5 mM</p><p><b>PiCa Mg2</b> = samples incubated in 1% FBS DMEM with Pi set to 3 mM, Ca²⁺ set to 2.4 mM and Mg²⁺ set to 2 mM</p><p><b>PiCa Mg5</b> = samples incubated in 1% FBS DMEM with Pi set to 3 mM, Ca²⁺ set to 2.4 mM and Mg²⁺ set to 5 mM</p><p>It is of note that these experiments were conducted concomitantly with and without cells. The table is indicating the presence of apatite in experiments with cells. Data of passive Ca / Pi deposition on MirrIR slides are discussed in a proper section.</p><p>Identification of the samples studied through µFTIR experiments.</p

Selected examples of infrared spectra from biopsies.

<p>a) Amorphous silica identified by a band at 1102 cm⁻¹, b) sodium hydrogen urate monohydrate identified by specific bands at 3600 and 1004 cm⁻¹, c) several calcium phosphates including whitlockite (peaks at 1080, 1025 cm⁻¹ and associated shoulders, d) octacalcium phosphate and carbapatite, identified by a shoulder at 1119 cm⁻¹, e) normal tissue, with signal of water (3300 cm⁻¹ and peaks around 1600 cm⁻¹) and proteins (peaks at 2900 cm⁻¹).</p

µFTIR maps obtained with calcium phosphate apatite features were collected for samples: PiCa, PiCa Mg2, PiCa Mg5, Pi4, Pi4 Mg2, Pi4 Mg5.

<p>The scale bar corresponds to 500 µm while the amplitude is between 0 and 10. Due to extended acquisition times, some maps were divided in 2 areas. The map size is 2000 µm x 4500 µm.</p

(A) SEM cartography of sample Pi4 Mg2 obtained with a magnification of 400 at 1 KeV (B) spatial distribution of Pi (purple maps), spatial distribution of Ca2+ (red maps) as obtained by EDX and SEM cartography (right pictures) obtained at 12 KeV for samples Pi4, Pi4 Mg2 and Pi4 Mg5.

<p>(A) SEM cartography of sample Pi4 Mg2 obtained with a magnification of 400 at 1 KeV (B) spatial distribution of Pi (purple maps), spatial distribution of Ca2+ (red maps) as obtained by EDX and SEM cartography (right pictures) obtained at 12 KeV for samples Pi4, Pi4 Mg2 and Pi4 Mg5.</p

Kidney Stone Classification Using Multimodal Multiphoton Microscopy

Kidney stones are a common form of nephrolithiasis, affecting up to 15% of the world’s population with a high probability of recurrence. These stones exhibit various chemical compositions and crystalline forms associated with different etiologies. Classification of the stones’ components is necessary to optimize treatment and suggest lifestyle changes to reduce the risk of recurrence. Current characterization methods usually require extensive sample preparation or are too detailed for the needs of a high-throughput laboratory. In this article, we present a kidney stone component classification scheme based on the multiphoton response of crushed samples that is label-free, requires minimal sample amounts, and simple preparation. We measure two-photon excited fluorescence, which is sensitive to protein content, second-harmonic generation, which is sensitive to crystalline symmetry, and polarization-resolved third-harmonic generation (pTHG), which is sensitive to crystal heterogeneity and birefringence. The combination of these three contrast modes can distinguish different materials, specifically calcium oxalate in monohydrate (COM), dihydrate (COD), or amorphous forms, cystine, and carbonate apatite. In addition, pTHG images have the potential to distinguish between COM and COD fragments and to provide information on the submicron organization of carbonate apatite fragments

Survival study and kidney injury in 129S2/SvPasCrl and C57BL/6J mice.

<p><b>A</b>, Survival proportions of mice from both strains before sacrifice at 16 weeks. 129S2/SvPasCrl mortality was significantly increased (p = 0.045) and necropsy revealed kidney dilation due to urolithiasis. <b>B</b>, Infrared cartography of 129S2/SvPasCrl kidney slices revealed the presence of focal cystine aggregates in renal tissues (cystine tubular casts). <b>C</b>, Renal function as assessed by enzymatic serum creatinine dosage was not significantly impaired in 129S1/SvPasCrl mice (n = 6) compared to C57BL/6J mice (n = 9) at 16 weeks (p = 0.11). <b>D–F</b>, Macrophage infiltrate in kidney tissues was assessed by morphometric analyses of F4-80 immunostaining and was increased in 129S2/SvPasCrl mice in comparison with C57BL/6J (n = 5/group, p = 0.046). <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0102700#pone-0102700-g003" target="_blank">Figures 3E</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0102700#pone-0102700-g003" target="_blank">3F</a> are representative of macrophage infiltrates in C57BL/6 and 129S1/SvPasCrl mice respectively (magnification x200+ zoom). <b>G</b>, Fibrosis assessed by sirius red morphometric analysis did not evidence significant fibrosis amount in both strains (percentage of fibrotic area, p = NS).</p