1. Endothelial cell injury induced by preservation solutions: a confocal microscopy study 
Francesco Alamanni, MD, Alessandro Parolari, MD, PhD, Rossana Visigalli, BS, PhD, Ovidio Bussolati, MD, PhD, Patrizia Rubini, MD, PhD, Roberto Sala, MD, PhD, Luigi Bonati, MD, Gian Carlo Gazzola, MD, Paolo Biglioli, MD, Valeria Dall’Asta, BS 
The Annals of Thoracic Surgery 
Volume 73, Issue 5, Pages (May 2002)
DOI: /S (02)

2. Fig 1
Two sets of cells were treated in parallel for each strain of human saphenous vein endothelial cells (HSVECs). At time 0, medium was substituted with normothermic culture medium (37°C M199), with hypothermic culture medium (4°C M199), or with the indicated hypothermic (4°C) storage solutions. After a 6-h treatment, cells were incubated in normothermic M199 up to 48 h. At the indicated times, cell number was determined as described in Material and Methods. Histograms are means of eight triplicate values each obtained in a distinct strain of HSVECs with SD.
The Annals of Thoracic Surgery  , DOI: ( /S (02) )

3. Fig 2
Two sets of cells were treated in parallel for each strain of human saphenous vein endothelial cells (HSVECs). At time 0, medium was substituted with normothermic culture medium (37°C M199), with hypothermic culture medium (4°C M199), or with the indicated hypothermic (4°C) storage solutions. After a 24-h treatment, cells were incubated in normothermic M199 up to 48 h. At the indicated times, cell number was determined as described in Material and Methods. Histograms are means of eight triplicate values each obtained in a distinct strain of HSVECs with SD.
The Annals of Thoracic Surgery  , DOI: ( /S (02) )

4. Fig 3
Confocal images of human saphenous vein endothelial cells (HSVECs) after 6 h of hypothermic preservation and 48 h of additional simulated reperfusion. The incubation medium of HSVECs was substituted for 6 h with normothermic (37°C) culture medium (M199; A and G), hypothermic (4°C) culture medium (M199; B and H), Celsior (C and I), Euro-Collins (D and J), St. Thomas II (E and K), or Wisconsin (F and L) solutions. Cell loading with calcein-AM and confocal observation of representative fields were performed at the end of the treatment (A-F) or after a subsequent 48-h incubation in warm M199 (G-L). The experiment, repeated three times, yielded comparable results. Intensities were expressed on a scale from black (zero) to white (maximal intensity, 255); images were rendered in a 256 RGB pseudocolors scale (L, inset) from black (zero), through different hues of blue, yellow and red, to white (255). (Bar = 20 μm.)
The Annals of Thoracic Surgery  , DOI: ( /S (02) )

5. Fig 4
Confocal images of human saphenous vein endothelial cells (HSVECs) after 5 h of hypothermic preservation. The incubation medium of cells was substituted for 5 h with normothermic culture medium (M199; A, control), hypothermic Euro-Collins (B), or hypothermic St. Thomas II (C) solutions. Cell loading with calcein-AM and confocal observation of representative fields were performed at the end of the treatment. (Bar = 10 μm.)
The Annals of Thoracic Surgery  , DOI: ( /S (02) )

6. Fig 5
Confocal images of human saphenous vein endothelial cells (HSVECs) after 24 h of hypothermic preservation and an additional 48 h of simulated reperfusion. The incubation medium of cells was substituted for 24 h with normothermic culture medium (M199; A and G), hypothermic culture medium (M199, B and H), Celsior (C and I), Euro-Collins (D and J), St. Thomas II (E and K), or Wisconsin (F and L) solutions. Cell loading with calcein-AM and confocal observation of representative fields were performed at the end of the treatment (A-F) or after a subsequent 48-h incubation in warm M199 (G-L). For other details, see the legend to Figure 3. (Bar = 20 μm.)
The Annals of Thoracic Surgery  , DOI: ( /S (02) )