Media 1: In vivo structural imaging of the cornea by polarization-resolved second harmonic microscopy

Originally published in Biomedical Optics Express on 01 January 2012 (boe-3-1-1

Media 4: In vivo structural imaging of the cornea by polarization-resolved second harmonic microscopy

Originally published in Biomedical Optics Express on 01 January 2012 (boe-3-1-1

Media 3: In vivo structural imaging of the cornea by polarization-resolved second harmonic microscopy

Originally published in Biomedical Optics Express on 01 January 2012 (boe-3-1-1

Media 2: In vivo structural imaging of the cornea by polarization-resolved second harmonic microscopy

Originally published in Biomedical Optics Express on 01 January 2012 (boe-3-1-1

Media 5: In vivo structural imaging of the cornea by polarization-resolved second harmonic microscopy

Originally published in Biomedical Optics Express on 01 January 2012 (boe-3-1-1

Confocal microscopy (CM) and SHG microscopy imaging of the DM from control and diabetic rat corneas.

<p>Unstained intact rat cornea observed by (a and c) <i>in vivo</i> CM and (b, d–i, j) <i>ex vivo</i> SHG microscopy detected in the (e) backward direction and in the (b,d,i–j) forward direction : (b-e) frontal optical sections, (i) 3D view and (j) transverse numerical reconstruction of the DM (image size: 108×108×7 µm³). Intensity profiles from (f) CM, (g) forward-detected and (h) backward-detected SHG microscopy along 100 µm are plotted under the corresponding images. The number of detected photons has been corrected from the channel sensitivity. St: stroma, DM: Descemet’s membrane. White arrows indicate collagen abnormal deposits in the DM. The look-up-table (LUT) used for SHG images is indicated near (b).</p

Methodology to compare the different imaging techniques on rat and human corneas.

<p>Cornea preparation, orientation of the different histological and numerical sections and imaging geometry are indicated for each imaging technique. Histological sections are unstained for SHG microscopy, stained with toluidine blue for transmitted light microscopy and stained with uranyl and lead citrate solutions for transmission electron microscopy.</p

Multimodal imaging of histological sections from the same diabetic rat cornea.

<p>(a–c) Frontal and (d–f) transverse histological sections of the cornea observed (a, d) by transmitted light microcopy, (b, c and e) by TEM, where insets show long-spacing collagen, and (f) by SHG microscopy. St: stroma, DM: Descemet’s membrane. White arrows indicate collagen abnormal deposits in the DM.</p

Human corneas: clinical data and observations by SHG microscopy and other imaging techniques.

<p>Grading is as follows: N/A: not available, −: absence of fibrillar collagen, +: few fibrillar collagen, ++: great concentration of fibrillar collagen, +++: fibrillar collagen over the whole field of view.</p

Diabetic and control rat corneas: clinical data and observations by SHG microscopy and other imaging techniques.

<p>Grading is as follows: N/A: not available, −: absence of fibrillar collagen, +: few fibrillar collagen, ++: great concentration of fibrillar collagen, +++: fibrillar collagen over the whole field of view.</p